Quinoline oxadiazoles as cytoprotective agents

ABSTRACT

Provided herein are compounds of Formula I, pharmaceutical compositions thereof, and methods of their use for treating, preventing, or ameliorating one or more symptoms of a neurological disease, neurodegenerative disorder, or diabetes.

1. CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation-in-part of U.S. patent applicationSer. No. 15/361,482, filed Nov. 27, 2016 (pending) which claims thebenefit of priority from U.S. Provisional Application No. 62/260,512,filed Nov. 28, 2015, each of which is incorporated by reference hereinin its entirety.

2. FIELD

Provided herein are novel compounds, pharmaceutical compositionsthereof, and methods of use for promoting neuroprotection andcytoprotection, and/or inhibiting cell death.

3. BACKGROUND

Effective drugs that are disease modifying remain elusive for thetreatment of neurodegenerative diseases like Alzheimer's disease (AD)and Parkinson's disease (PD), and diabetes including type I and type II.Majority of the approved medicines for these conditions treat thesymptoms rather than targeting slowing or halting disease progression.Beta cell death is an accepted major causal feature of both diabetes.Cnop et al., Diabetes 2005, 54, S97-107. Neuron death is also the majorcause of cognitive decline in Alzheimer's disease. Niikura et al., Curr.Neuropharmacol. 2006, 4, 139-147. Neuron death is also implicated inParkinson's and Huntington's diseases. Anglade et al., Histol.Histopathol. 1997, 12, 25-31.

A major cause of the cell death seen in diabetes and neurodegenerativedisorders is a condition known as endoplasmic reticulum (ER) stress.Ozcan et al., Annu. Rev. Med. 2012, 63, 317-328. This ER stress iscaused by abnormal calcium homeostasis, often caused by compromisedfunction of the ER calcium pump, sarco/endoplasmic reticulum calciumATPase (SERCA). Krebs et al., Biochem. Biophys. Res. Commun. 2015, 460,114-121. Thus, an alternative approach to the treatment ofneurodegeneration and diabetes is to target aberrant calcium signalingcascades that prove to be pathogenic. Stabilization of calcium signalingtargets a pathogenic mechanism that is tied to many major features andrisk factors of neurodegenerative diseases and diabetes. Rather thantargeting a single endpoint this strategy aims to normalize calciumdyshomeostasis that has been recognized as a pathogenic accelerant inthese diseases. Wang et al., J Cell Biol. 2012, 197, 857-867.

The ER is an organelle, which plays an essential role in multiplecellular processes that are central for cell survival and normalcellular functions. Those vital cellular processes include intracellularcalcium homeostasis, protein secretion, and lipid biosynthesis. Anelliet al., EMBO J. 2008, 27, 315-327; Pizzo et al., Trends Cell Biol. 2007,17, 511-517; Ma et al., J. Chem. Neuroanat. 2004, 28, 51-65.

Proper ER function and cellular health is dependent on the maintenanceof a precise intracellular calcium balance of low cytosolic Ca²⁺ ionconcentration coupled with high ER Ca²⁺ ion concentration. La Rovere, etal., Cell Calcium 2016, 60, 74-87; Rizzuto, et al. Nat. Rev. Mol. CellBiol. 2012, 13, 566-578; Reddish, et al. Int. J. Mol. Sci. 2017, 18,1024. Numerous diseases are associated with aberrant calcium homeostasisand transport. Mekahli, et al., Cold Spring Harb. Perspect. Biol. 2011,3, a004317.

Perturbation of ER homeostasis leads to accumulation of unfolded proteinin the ER, triggering an evolutionarily conserved response known as theunfolded protein response (UPR). Ron et al., Nat. Rev. Mol. Cell Biol.2007, 8, 519-529; Malhotra et al., Semin. Cell Dev. Biol. 2007, 18,716-731. Disturbances that lead to ER stress include, for example,disturbances in cellular redox regulation, glucose deprivation,aberration of calcium regulation in the ER, viral infection, high-fatdiet, protein-inclusion-body diseases (e.g., chronic neurodegenerativediseases), and inclusion-body myositis. Kim et al., Nat. Rev. Drug Dis.2008, 7, 1013-1030; Ma et al., J. Chem. Neuroanat. 2004, 28, 51-65;Ozcan et al., Science 2004, 306, 457-461; Frand et al., Trends CellBiol. 2000, 10, 203-3 10. ER stress has been linked to a wide range ofdiseases, including neurodegeneration (e.g., Alzheimer's disease,Parkinson's disease, amyotrophic lateral sclerosis, polyglutaminedisease, and prion disease), stroke, bipolar disorder, heart disease,atherosclerosis, cancer, diabetes (types 1 and 2), muscle degeneration,inflammatory diseases, and autoimmune disease. Kim et al., Nat. Rev.Drug Dis. 2008, 7, 1013-1030; Oyadomari et al., Cell Death Differ. 2004,11, 381-389.

Sarcoplasmic/endoplasmic reticulum Ca²⁺ ATPase (SERCA), is a majorregulator of ER stress and glucose homeostasis in obesity. Park et al.,Proc. Natl. Acad. Sci. U.S.A. 2010, 107, 19320-19325. Obesity disruptsintracellular Ca²⁺ homeostasis and induces ER stress. Fu et al., Nature2011, 473, 528-531. Chronic activation of ER stress has been implicatedin the development of insulin resistance and diabetes in obesity.Hotamisligil, Cell 2010, 140, 900-917; Kim et al., Nat. Rev. DrugDiscov. 2008, 7, 1013-1030. Restoration of SERCA expression in the liverof diabetic mice either via gene therapy or pharmacological activationreduced ER stress and improved the diabetic phenotype. Kang, et al., J.Biol. Chem. 2016, 291, 5185-5198; Park, et al., Proc. Natl. Acad. Sci.U.S.A. 2010, 107, 19320-19325. Restoration of Ca²⁺ homeostasis via SERCAactivation has been shown to alleviate dyskinesia in a model ofParkinson's disease. Dahl, Bioorg. Med. Chem. 2017, 25, 53-57. SERCAactivation has also been shown to improve memory and coordination in atransgenic mouse model of Alzheimer's disease. Krajnak & Dahl, Bioorg.Med. Chem Lett. 2018, 28, 1591-1594. Therefore, there is a need fortherapeutic agents capable of reducing ER stress or restoring ER calciumhomeostasis for treating diabetes and neurodegeneration.

4. SUMMARY

This disclosure provides compositions and methods comprising compoundsof Formula I.

or an enantiomer, a mixture of enantiomers, a mixture of two or morediastereomers, or an isotopic variant thereof; or a pharmaceuticallyacceptable salt, solvate, or hydrate thereof which are useful inpromoting cytoprotection and/or inhibiting cell death. The inventionalso comprises pharmaceutical compositions comprising a therapeuticallyeffective amount of compound of Formula I, or a pharmaceuticallyacceptable salt, solvate, or hydrate thereof. The invention disclosedherein is also directed to a method of promoting the generation orsurvival of beta cells or neurons in a patient in need thereof indiabetes or neurodegenerative and related diseases.

5. DETAILED DESCRIPTION 5.1. Definitions

To facilitate understanding of the disclosure set forth herein, a numberof terms are defined below.

Generally, the nomenclature used herein and the laboratory procedures inorganic chemistry, medicinal chemistry, and pharmacology describedherein are those well known and commonly employed in the art. Unlessdefined otherwise, all technical and scientific terms used hereingenerally have the same meaning as commonly understood by one ofordinary skill in the art to which this disclosure belongs.

The term “subject” refers to an animal, including, but not limited to, aprimate (e.g., human), cow, pig, sheep, goat, horse, dog, cat, rabbit,rat, or mouse. The terms “subject” and “patient” are usedinterchangeably herein in reference, for example, to a mammaliansubject. In one embodiment, the subject is a human.

The terms “treat,” “treating,” and “treatment” are meant to includealleviating or abrogating a disorder, disease, or condition, or one ormore symptoms of the disorder, disease, or condition; or alleviating oreradicating the cause(s) of the disorder, disease, or condition itself.

The terms “prevent,” “preventing,” and “prevention” are meant to includea method of delaying and/or precluding the onset of a disorder, disease,or condition, and/or its attendant symptoms; barring a subject fromacquiring a disorder, disease, or condition; or reducing a subject'srisk of acquiring a disorder, disease, or condition.

The term “therapeutically effective amount” is meant to include theamount of a compound that, when administered, is sufficient to preventdevelopment of, or alleviate to some extent, one or more symptoms of thedisorder, disease, or condition being treated. The term “therapeuticallyeffective amount” also refers to the amount of a compound that issufficient to elicit the biological or medical response of a biologicalmolecule (e.g., a protein, enzyme, RNA, or DNA), cell, tissue, system,animal, or human, which is being sought by a researcher, veterinarian,medical doctor, or clinician.

The term “pharmaceutically acceptable carrier,” “pharmaceuticallyacceptable excipient,” “physiologically acceptable carrier,” or“physiologically acceptable excipient” refers to a pharmaceuticallyacceptable material, composition, or vehicle, such as a liquid or solidfiller, diluent, solvent, or encapsulating material. In one embodiment,each component is “pharmaceutically acceptable” in the sense of beingcompatible with other ingredients of a pharmaceutical formulation, andsuitable for use in contact with the tissue or organ of humans andanimals without excessive toxicity, irritation, allergic response,immunogenicity, or other problems or complications, commensurate with areasonable benefit/risk ratio. See, Remington: The Science and Practiceof Pharmacy, 21st Edition, Lippincott Williams & Wilkins: Philadelphia,Pa., 2005; Handbook of Pharmaceutical Excipients, 7th Edition, Rowe etal., Eds., The Pharmaceutical Press and the American PharmaceuticalAssociation: 2012; Handbook of Pharmaceutical Additives, 3rd Edition,Ash and Ash Eds., Gower Publishing Company: 2007; and PharmaceuticalPreformulation and Formulation, 2nd Edition, Gibson Ed., CRC Press LLC:Boca Raton, Fla., 2009.

The term “about” or “approximately” means an acceptable error for aparticular value as determined by one of ordinary skill in the art,which depends in part on how the value is measured or determined. Incertain embodiments, the term “about” or “approximately” means within 1,2, 3, or 4 standard deviations. In certain embodiments, the term “about”or “approximately” means within 50%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%,4%, 3%, 2%, 1%, 0.5%, or 0.05% of a given value or range.

The terms “active ingredient” and “active substance” refer to acompound, which is administered, alone or in combination with one ormore pharmaceutically acceptable excipients, to a subject for treating,preventing, or ameliorating one or more symptoms of a disorder, disease,or condition. As used herein, “active ingredient” and “active substance”may be an optically active isomer of a compound described herein.

The terms “drug,” “therapeutic agent,” and “chemotherapeutic agent”refer to a compound or a pharmaceutical composition thereof, which isadministered to a subject for treating, preventing, or ameliorating oneor more symptoms of a disorder, disease, or condition.

The term “endoplasmic reticulum stress” or “ER stress” refers toperturbation of endoplasmic reticulum homeostasis, e.g., perturbation ofthe protein folding functionality of the endoplasmic reticulum.

The term “naturally occurring” or “native” when used in connection witha biological material, such as a nucleic acid (e.g., a DNA or RNA), apolypeptide, and a host cell, refers to a material which is found innature and is not manipulated by man. Similarly, “non-naturallyoccurring” or “non-native” refers to a material that is not found innature or that has been structurally modified or synthesized by man.

The term “SERCA” or “sarco(endo)plasmic reticulum Ca²⁺ ATPase” refers toa sarcoplasmic/endoplasmic reticulum Ca²⁺ ATPase or a variant thereof.The term “SERCA variant” is intended to include proteins substantiallyhomologous to a native SERCA, i.e., proteins having one or morenaturally or non-naturally occurring amino acid deletions, insertions,or substitutions (e.g., SERCA derivatives, homologs, and fragments), ascompared to the amino acid sequence of a native SERCA. The amino acidsequence of a SERCA variant is at least about 80% identical, at leastabout 90% identical, or at least about 95% identical to a native SERCA.SERCA enzymes are classified into at least three classes: SERCA1,SERCA2, and SERCA3. Stutzmann et al., Pharmacol. Rev. 2011, 63, 700-727;Andersen et al., Acta Physiol. Scand. Suppl. 1998, 643, 45-54. Class Iincludes SERCA1a and SERCA1b. Class II includes SERCA2a and SERCA2b.Class III includes SERCA3a, SERCA3b, and SERCA3c.

The terms “SERCA-mediated disorder, disease, or condition” and “adisorder, disease, or condition mediated by SERCA” refer to a disorder,disease, or condition in which modulation of a SERCA activity results insome effect on the underlying disorder, disease, or condition, e.g., aSERCA agonist results in some improvement in at least some of patientsbeing treated.

The term “alkyl” refers to a linear or branched saturated monovalenthydrocarbon radical, wherein the alkyl is optionally substituted withone or more substituents Q as described herein. The term “alkyl” alsoencompasses both linear and branched alkyl, unless otherwise specified.In certain embodiments, the alkyl is a linear saturated monovalenthydrocarbon radical that has 1 to 20 (C₁₋₂₀), 1 to 15 (C₁₋₁₅), 1 to 10(C₁₋₁₀), or 1 to 6 (C₁₋₆) carbon atoms, or branched saturated monovalenthydrocarbon radical of 3 to 20 (C₃₋₂₀), 3 to 15 (C₃₋₁₅), 3 to 10(C₃₋₁₀), or 3 to 6 (C₃₋₆) carbon atoms. As used herein, linear C₁₋₆ andbranched C₃₋₆ alkyl groups are also referred as “lower alkyl.” Examplesof alkyl groups include, but are not limited to, methyl, ethyl, propyl(including all isomeric forms), n-propyl, isopropyl, butyl (includingall isomeric forms), n-butyl, isobutyl, sec-butyl, t-butyl, pentyl(including all isomeric forms), and hexyl (including all isomericforms). For example, C₁₋₆ alkyl refers to a linear saturated monovalenthydrocarbon radical of 1 to 6 carbon atoms or a branched saturatedmonovalent hydrocarbon radical of 3 to 6 carbon atoms.

The term “alkenyl” refers to a linear or branched monovalent hydrocarbonradical, which contains one or more, in one embodiment, one, two, three,four, or five, in another embodiment, one, carbon-carbon double bond(s).In certain embodiments, the alkenyl is optionally substituted with oneor more substituents Q as described herein. The term “alkenyl” alsoembraces radicals having “cis” and “trans” configurations, oralternatively, “Z” and “E” configurations, as appreciated by those ofordinary skill in the art. As used herein, the term “alkenyl”encompasses both linear and branched alkenyl, unless otherwisespecified. For example, C₂₋₆ alkenyl refers to a linear unsaturatedmonovalent hydrocarbon radical of 2 to 6 carbon atoms or a branchedunsaturated monovalent hydrocarbon radical of 3 to 6 carbon atoms. Incertain embodiments, the alkenyl is a linear monovalent hydrocarbonradical of 2 to 20 (C₂₋₂₀), 2 to 15 (C₂₋₁₅), 2 to 10 (C₂₋₁₀), or 2 to 6(C₂₋₆) carbon atoms, or a branched monovalent hydrocarbon radical of 3to 20 (C₃₋₂₀), 3 to 15 (C₃₋₁₅), 3 to 10 (C₃₋₁₀), or 3 to 6 (C₃₋₆) carbonatoms. Examples of alkenyl groups include, but are not limited to,ethenyl, propen-1-yl, propen-2-yl, allyl, butenyl, and 4-methylbutenyl.

The term “alkynyl” refers to a linear or branched monovalent hydrocarbonradical, which contains one or more, in one embodiment, one, two, three,four, or five, in another embodiment, one, carbon-carbon triple bond(s).In certain embodiments, the alkynyl is optionally substituted with oneor more substituents Q as described herein. The term “alkynyl” alsoencompasses both linear and branched alkynyl, unless otherwisespecified. In certain embodiments, the alkynyl is a linear monovalenthydrocarbon radical of 2 to 20 (C₂₋₂₀), 2 to 15 (C₂₋₁₅), 2 to 10(C₂₋₁₀), or 2 to 6 (C₂₋₆) carbon atoms, or a branched monovalenthydrocarbon radical of 3 to 20 (C₃₋₂₀), 3 to 15 (C₃₋₁₅), 3 to 10(C₃-10), or 3 to 6 (C₃₋₆) carbon atoms. Examples of alkynyl groupsinclude, but are not limited to, ethynyl (—C≡CH) and propargyl(—CH₂C≡CH). For example, C₂₋₆ alkynyl refers to a linear unsaturatedmonovalent hydrocarbon radical of 2 to 6 carbon atoms or a branchedunsaturated monovalent hydrocarbon radical of 3 to 6 carbon atoms.

The term “cycloalkyl” refers to a cyclic saturated or non-aromaticunsaturated, bridged or non-bridged monovalent hydrocarbon radical,which is optionally substituted with one or more substituents Q asdescribed herein. In certain embodiments, the cycloalkyl is a cyclicsaturated bridged or non-bridged monovalent hydrocarbon radical. Incertain embodiments, the cycloalkyl has from 3 to 20 (C₃₋₂₀), from 3 to15 (C₃₋₁₅), from 3 to 10 (C₃₋₁₀), or from 3 to 7 (C₃₋₇) carbon atoms.Examples of cycloalkyl groups include, but are not limited to,cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl,bicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptyl, decalinyl, and adamantyl.

The term “aryl” refers to a monocyclic aromatic group and/or multicyclicmonovalent aromatic group that contain at least one aromatic hydrocarbonring. In certain embodiments, the aryl has from 6 to 20 (C₆₋₂₀), from 6to 15 (C₆₋₁₅), or from 6 to 10 (C₆₋₁₀) ring atoms. Examples of arylgroups include, but are not limited to, phenyl, naphthyl, fluorenyl,azulenyl, anthryl, phenanthryl, pyrenyl, biphenyl, and terphenyl. Incertain embodiments, the term “aryl” refers to a bicyclic or tricycliccarbon ring, where one of the rings is aromatic and the others of whichmay be saturated, partially unsaturated, or aromatic, for example,dihydronaphthyl, indenyl, indanyl, or tetrahydronaphthyl (tetralinyl).In certain embodiments, the aryl is optionally substituted with one ormore substituents Q as described herein.

The term “aralkyl” or “arylalkyl” refers to a monovalent alkyl groupsubstituted with one or more aryl groups. In certain embodiments, thearalkyl has from 7 to 30 (C₇₋₃₀), from 7 to 20 (C₇₋₂₀), or from 7 to 16(C₇₋₁₆) carbon atoms. Examples of aralkyl groups include, but are notlimited to, benzyl, 1-phenylethyl, 2-phenylethyl, and 3-phenylpropyl. Incertain embodiments, the aralkyl is optionally substituted with one ormore substituents Q as described herein.

The term “heteroaryl” refers to a monovalent monocyclic aromatic groupor monovalent polycyclic aromatic group that contain at least onearomatic ring, wherein at least one aromatic ring contains one or moreheteroatoms, each of which is independently selected from O, S, N, andP, in the ring. A heteroaryl group is bonded to the rest of a moleculethrough its aromatic ring. Each ring of a heteroaryl group can containone or two O atoms, one or two S atoms, one to four N atoms, and/or oneor two P atoms, provided that the total number of heteroatoms in eachring is four or less and each ring contains at least one carbon atom. Incertain embodiments, the heteroaryl has from 5 to 20, from 5 to 15, orfrom 5 to 10 ring atoms. Examples of monocyclic heteroaryl groupsinclude, but are not limited to, furanyl, imidazolyl, isothiazolyl,isoxazolyl, oxadiazolyl, oxadiazolyl, oxazolyl, pyrazinyl, pyrazolyl,pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, thiadiazolyl, thiazolyl,thienyl, tetrazolyl, triazinyl, and triazolyl. Examples of bicyclicheteroaryl groups include, but are not limited to, benzofuranyl,benzimidazolyl, benzoisoxazolyl, benzopyranyl, benzothiadiazolyl,benzothiazolyl, benzothienyl, benzotriazolyl, benzoxazolyl, furopyridyl,imidazopyridinyl, imidazothiazolyl, indolizinyl, indolyl, indazolyl,isobenzofuranyl, isobenzothienyl, isoindolyl, isoquinolinyl,isothiazolyl, naphthyridinyl, oxazolopyridinyl, phthalazinyl,pteridinyl, purinyl, pyridopyridyl, pyrrolopyridyl, quinolinyl,quinoxalinyl, quinazolinyl, thiadiazolopyrimidyl, and thienopyridyl.Examples of tricyclic heteroaryl groups include, but are not limited to,acridinyl, benzindolyl, carbazolyl, dibenzofuranyl, perimidinyl,phenanthrolinyl, phenanthridinyl, phenarsazinyl, phenazinyl,phenothiazinyl, phenoxazinyl, and xanthenyl. In certain embodiments, theheteroaryl is optionally substituted with one or more substituents Q asdescribed herein.

The term “heterocyclyl” or “heterocyclic” refers to a monovalentmonocyclic non-aromatic ring system or monovalent polycyclic ring systemthat contains at least one non-aromatic ring, wherein one or more of thenon-aromatic ring atoms are heteroatoms, each of which is independentlyselected from O, S, N, and P; and the remaining ring atoms are carbonatoms. In certain embodiments, the heterocyclyl or heterocyclic grouphas from 3 to 20, from 3 to 15, from 3 to 10, from 3 to 8, from 4 to 7,or from 5 to 6 ring atoms. A heterocyclyl group is bonded to the rest ofa molecule through its non-aromatic ring. In certain embodiments, theheterocyclyl is a monocyclic, bicyclic, tricyclic, or tetracyclic ringsystem, which may be spiro, fused, or bridged, and in which nitrogen orsulfur atoms may be optionally oxidized, nitrogen atoms may beoptionally quaternized, and some rings may be partially or fullysaturated, or aromatic. The heterocyclyl may be attached to the mainstructure at any heteroatom or carbon atom which results in the creationof a stable compound. Examples of heterocyclic groups include, but arenot limited to, azepinyl, benzodioxanyl, benzodioxolyl, benzofuranonyl,benzopyranonyl, benzopyranyl, benzotetrahydrofuranyl,benzotetrahydrothienyl, benzothiopyranyl, benzoxazinyl, β-carbolinyl,chromanyl, chromonyl, cinnolinyl, coumarinyl, decahydroisoquinolinyl,dihydrobenzisothiazinyl, dihydrobenzisoxazinyl, dihydrofuryl,dihydroisoindolyl, dihydropyranyl, dihydropyrazolyl, dihydropyrazinyl,dihydropyridinyl, dihydropyrimidinyl, dihydropyrrolyl, dioxolanyl,1,4-dithianyl, furanonyl, imidazolidinyl, imidazolinyl, indolinyl,isobenzotetrahydrofuranyl, isobenzotetrahydrothienyl, isochromanyl,isocoumarinyl, isoindolinyl, isothiazolidinyl, isoxazolidinyl,morpholinyl, octahydroindolyl, octahydroisoindolyl, oxazolidinonyl,oxazolidinyl, oxiranyl, piperazinyl, piperidinyl, 4-piperidonyl,pyrazolidinyl, pyrazolinyl, pyrrolidinyl, pyrrolinyl, quinuclidinyl,tetrahydrofuryl, tetrahydroisoquinolinyl, tetrahydropyranyl,tetrahydrothienyl, thiamorpholinyl, thiazolidinyl, tetrahydroquinolinyl,and 1,3,5-trithianyl. In certain embodiments, the heterocyclyl isoptionally substituted with one or more substituents Q as describedherein.

The term “halogen”, “halide” or “halo” refers to fluorine, chlorine,bromine, and/or iodine.

The term “optionally substituted” is intended to mean that a group orsubstituent, such as an alkyl, alkenyl, alkynyl, cycloalkyl, aryl,aralkyl, heteroaryl, and heterocyclyl group, may be substituted with oneor more substituents Q, each of which is independently selected from,e.g., (a) oxo (═O), cyano (—CN), halo, and nitro (—NO₂); (b) C₁₋₆ alkyl,C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₁₀ cycloalkyl, C₆₋₁₄ aryl, C₇₋₁₅ aralkyl,heteroaryl, and heterocyclyl, each of which is further optionallysubstituted with one or more, in one embodiment, one, two, three, four,or five, substituents Q^(a); and (c) —C(O)R^(a), —C(O)OR^(a),—C(O)NR^(b)R^(c), —C(NR^(a))NR^(b)R^(c), —OR^(a), —OC(O)R^(a),—OC(O)OR^(a), —OC(O)NR^(b)R^(c), —OC(═NR^(a))NR^(b)R^(c), —OS(O)R^(a),—OS(O)₂R^(a), —OS(O)NR^(b)R^(c), —OS(O)₂NR^(b)R^(c), —NR^(b)R^(c),—NR^(a)C(O)R^(d), —NR^(a)C(O)OR^(d), —NR^(a)C(O)NR^(b)R^(c),—NR^(a)C(═NR^(d))NR^(b)R^(c), —NR^(a)S(O)R^(d), —NR^(a)S(O)₂R^(d),—NR^(a)S(O)NR^(b)R^(c), —NR^(a)S(O)₂NR^(b)R^(c), —P(O)R^(a)R^(d),—P(O)(OR^(a))R^(d), —P(O)(OR^(a))(OR^(d)), —SR^(a), —S(O)R^(a),—S(O)₂R^(a), —S(O)NR^(b)R^(c), and —S(O)₂NR^(b)R^(c), wherein eachR^(a), R^(b), R^(c), and R^(d) is independently (i) hydrogen; (ii) C₁₋₆alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₁₀ cycloalkyl, C₆₋₁₄ aryl, C₇₋₁₅aralkyl, heteroaryl, or heterocyclyl, each of which is optionallysubstituted with one or more, in one embodiment, one, two, three, orfour, substituents Q^(a); or (iii) R^(b) and R^(c) together with the Natom to which they are attached form heteroaryl or heterocyclyl, each ofwhich is optionally substituted with one or more, in one embodiment,one, two, three, or four, substituents Q^(a). As used herein, all groupsdescribed herein that can be substituted are “optionally substituted,”unless otherwise specified.

In one embodiment, each substituent Q^(a) is independently selected fromthe group consisting of (a) oxo, cyano, halo, and nitro; and (b) C₁₋₆alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₁₀ cycloalkyl, C₆₋₁₄ aryl, C₇₋₁₅aralkyl, heteroaryl, and heterocyclyl; and (c) —C(O)R^(e), —C(O)OR^(e),—C(O)NR^(f)R^(g), —C(NR^(e))NR^(f)R^(g), —OR^(e), —OC(O)R^(e),—OC(O)OR^(e), —OC(O)NR^(f)R^(g), —OC(═NR^(e))NR^(f)R^(g), —OS(O)R^(e),—OS(O)₂R^(e), —OS(O)NR^(f)R^(g), —OS(O)₂NR^(f)R^(g), —NR^(f)R^(g),—NR^(e)C(O)R^(h), —NR^(e)C(O)OR^(h), —NR^(e)C(O)NR^(f)R^(g),—NR^(e)C(═NR^(h))NR^(f)R^(g), —NR^(e)S(O)R^(h), —NR^(e)S(O)₂R^(h),—NR^(e)S(O)NR^(f)R^(g), —NR^(e)S(O)₂NR^(f)R^(g), —P(O)R^(e)R^(h),—P(O)(OR^(e))R^(h), —P(O)(OR^(e))(OR^(h)), —SR^(e), —S(O)R^(e),—S(O)₂R^(e), —S(O)NR^(f)R^(g), and —S(O)₂NR^(f)R^(g); wherein eachR^(e), R^(f), R^(g), and R^(h) is independently (i) hydrogen, C₁₋₆alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₁₀ cycloalkyl, C₆₋₁₄ aryl, C₇₋₁₅aralkyl, heteroaryl, or heterocyclyl; or (ii) R^(f) and R^(g) togetherwith the N atom to which they are attached form heteroaryl orheterocyclyl.

In certain embodiments, “optically active” and “enantiomerically active”refer to a collection of molecules, which has an enantiomeric excess ofno less than about 50%, no less than about 70%, no less than about 80%,no less than about 90%, no less than about 91%, no less than about 92%,no less than about 93%, no less than about 94%, no less than about 95%,no less than about 96%, no less than about 97%, no less than about 98%,no less than about 99%, no less than about 99.5%, or no less than about99.8%. In certain embodiments, the compound comprises about 95% or moreof the desired enantiomer and about 5% or less of the less preferredenantiomer based on the total weight of the two enantiomers in question.

In describing an optically active compound, the prefixes R and S areused to denote the absolute configuration of the optically activecompound about its chiral center(s). The (+) and (−) are used to denotethe optical rotation of an optically active compound, that is, thedirection in which a plane of polarized light is rotated by theoptically active compound. The (−) prefix indicates that an opticallyactive compound is levorotatory, that is, the compound rotates the planeof polarized light to the left or counterclockwise. The (+) prefixindicates that an optically active compound is dextrorotatory, that is,the compound rotates the plane of polarized light to the right orclockwise. However, the sign of optical rotation, (+) and (−), is notrelated to the absolute configuration of a compound, R and S.

The term “isotopic variant” refers to a compound that contains anunnatural proportion of an isotope at one or more of the atoms thatconstitute such a compound. In certain embodiments, an “isotopicvariant” of a compound contains unnatural proportions of one or moreisotopes, including, but not limited to, hydrogen (¹H), deuterium (²H),tritium (³H), carbon-11 (¹¹C), carbon-12 (¹²C), carbon-13 (¹³C),carbon-14 (¹⁴C), nitrogen-13 (¹³N), nitrogen-14 (¹⁴N), nitrogen-15(¹⁵N), oxygen-14 (¹⁴O), oxygen-15 (¹⁵O), oxygen-16 (¹⁶O), oxygen-17(¹⁷O), oxygen-18 (¹⁸O), fluorine-17 (¹⁷F), fluorine-18 (¹⁸F),phosphorus-31 (³¹P), phosphorus-32 (³²P), phosphorus-33 (³³P), sulfur-32(³²S), sulfur-33 (³³S), sulfur-34 (³⁴S), sulfur-35 (³⁵S), sulfur-36(³⁶S), chlorine-35 (³⁵Cl), chlorine-36 (³⁶Cl), chlorine-37 (³⁷Cl),bromine-79 (⁷⁹Br), bromine-81 (⁸¹Br), iodine-123 (¹²³I), iodine-125(¹²⁵I), iodine-127 (¹²⁷I), iodine-129 (¹²⁹I), and iodine-131 (¹³¹I). Incertain embodiments, an “isotopic variant” of a compound is in a stableform, that is, non-radioactive. In certain embodiments, an “isotopicvariant” of a compound contains unnatural proportions of one or moreisotopes, including, but not limited to, hydrogen (¹H), deuterium (²H),carbon-12 (¹²C), carbon-13 (¹³C), nitrogen-14 (¹⁴N), nitrogen-15 (¹⁵N),oxygen-16 (¹⁶O), oxygen-17 (¹⁷O), oxygen-18 (¹⁸O), fluorine-17 (¹⁷F),phosphorus-31 (³¹P), sulfur-32 (³²S), sulfur-33 (³³S), sulfur-34 (³⁴S),sulfur-36 (³⁶S), chlorine-35 (³⁵Cl), chlorine-37 (³⁷Cl), bromine-79(⁷⁹Br), bromine-81 (⁸¹Br), and iodine-127 (¹²⁷I). In certainembodiments, an “isotopic variant” of a compound is in an unstable form,that is, radioactive. In certain embodiments, an “isotopic variant” of acompound contains unnatural proportions of one or more isotopes,including, but not limited to, tritium (³H), carbon-11 (¹¹C), carbon-14(¹⁴C), nitrogen-13 (¹³N), oxygen-14 (¹⁴O), oxygen-15 (¹⁵O), fluorine-18(¹⁸F), phosphorus-32 (³²P), phosphorus-33 (³³P), sulfur-35 (³⁵S),chlorine-36 (³⁶Cl), iodine-123 (¹²³I), iodine-125 (¹²⁵I) iodine-129(¹²⁹I), and iodine-131 (¹³¹I). It will be understood that, in a compoundas provided herein, any hydrogen can be ²H, for example, or any carboncan be ¹³C, for example, or any nitrogen can be ¹⁵N, for example, or anyoxygen can be ¹⁸O, for example, where feasible according to the judgmentof one of skill. In certain embodiments, an “isotopic variant” of acompound contains unnatural proportions of deuterium (D).

The term “solvate” refers to a complex or aggregate formed by one ormore molecules of a solute, e.g., a compound provided herein, and one ormore molecules of a solvent, which present in a stoichiometric ornon-stoichiometric amount. Suitable solvents include, but are notlimited to, water, methanol, ethanol, n-propanol, isopropanol, andacetic acid. In certain embodiments, the solvent is pharmaceuticallyacceptable. In one embodiment, the complex or aggregate is in acrystalline form. In another embodiment, the complex or aggregate is ina noncrystalline form. Where the solvent is water, the solvate is ahydrate. Examples of hydrates include, but are not limited to, ahemihydrate, monohydrate, dihydrate, trihydrate, tetrahydrate, andpentahydrate.

The phrase “an enantiomer, a mixture of enantiomers, a mixture of two ormore diastereomers, or an isotopic variant thereof; or apharmaceutically acceptable salt, solvate, or hydrate thereof” has thesame meaning as the phrase “(i) an enantiomer, a mixture of enantiomers,a mixture of two or more diastereomers, or an isotopic variant of thecompound referenced therein; (ii) a pharmaceutically acceptable salt,solvate, or hydrate of the compound referenced therein; or (iii) apharmaceutically acceptable salt, solvate, or hydrate of an enantiomer,a mixture of enantiomers, a mixture of two or more diastereomers, or anisotopic variant of the compound referenced therein.”

5.2. Compounds

Provided herein is a compound of Formula I:

or an enantiomer, a mixture of enantiomers, a mixture of two or morediastereomers, or an isotopic variant thereof; or a pharmaceuticallyacceptable salt, solvate, or hydrate thereof; wherein:

R² is methyl;

R³, R⁴, R⁵, R⁶, and R⁷ are each independently (a) hydrogen, cyano, orhalo or (b) C₁₋₆ alkyl, C₃₋₁₀ cycloalkyl, or heterocyclyl with theproviso that R³ and R⁷ are not concurrently Cl; and

R¹ is aryl or heteroaryl.

In one embodiment, provided herein is a compound selected from the groupconsisting of:

and isotopic variants thereof; and pharmaceutically acceptable salts,solvates, and hydrates thereof.

In certain embodiments, the compounds provided herein show activity asagonists of a SERCA. In certain embodiments, the compounds providedherein show activity as allosteric SERCA modulators. In certainembodiments, the compounds provided herein show activity as agonists ofa SERCA2b. In certain embodiments, the compounds provided herein showactivity as allosteric SERCA2b modulators.

In certain embodiments, the compounds provided herein show activity inreducing ER stress. In certain embodiments, the compounds providedherein show activity in increasing the Ca²⁺ concentration of an ER.

The compounds provided herein are intended to encompass all possiblestereoisomers, unless a particular stereochemistry is specified. Wherethe compound provided herein contains an alkenyl or alkenylene group,the compound may exist as one or mixture of geometric cis/trans (or Z/E)isomers. Where structural isomers are interconvertible, the compound mayexist as a single tautomer or a mixture of tautomers. This can take theform of proton tautomerism in the compound that contains, for example,an imino, keto, or oxime group; or so-called valence tautomerism in thecompound that contain an aromatic moiety. It follows that a singlecompound may exhibit more than one type of isomerism.

The compounds provided herein may be enantiomerically pure, such as asingle enantiomer or a single diastereomer, or be stereoisomericmixtures, such as a mixture of enantiomers, e.g., a racemic mixture oftwo enantiomers; or a mixture of two or more diastereomers. As such, oneof skill in the art will recognize that administration of a compound inits (R) form is equivalent, for compounds that undergo epimerization invivo, to administration of the compound in its (S) form. Conventionaltechniques for the preparation/isolation of individual enantiomersinclude synthesis from a suitable optically pure precursor, asymmetricsynthesis from achiral starting materials, or resolution of anenantiomeric mixture, for example, chiral chromatography,recrystallization, resolution, diastereomeric salt formation, orderivatization into diastereomeric adducts followed by separation.

When a compound provided herein contains an acidic or basic moiety, itmay also be provided as a pharmaceutically acceptable salt (See, Bergeet al., J. Pharm. Sci. 1977, 66, 1-19; and “Handbook of PharmaceuticalSalts, Properties, and Use,” Stahl and Wermuth, Ed.; Wiley-VCH and VHCA,Zurich, 2002).

Suitable acids for use in the preparation of pharmaceutically acceptablesalts include, but are not limited to, acetic acid, 2,2-dichloroaceticacid, acylated amino acids, adipic acid, alginic acid, ascorbic acid,L-aspartic acid, benzenesulfonic acid, benzoic acid, 4-acetamidobenzoicacid, boric acid, (+)-camphoric acid, camphorsulfonic acid,(+)-(1S)-camphor-10-sulfonic acid, capric acid, caproic acid, caprylicacid, cinnamic acid, citric acid, cyclamic acid, cyclohexanesulfamicacid, dodecylsulfuric acid, ethane-1,2-disulfonic acid, ethanesulfonicacid, 2-hydroxy-ethanesulfonic acid, formic acid, fumaric acid,galactaric acid, gentisic acid, glucoheptonic acid, D-gluconic acid,D-glucuronic acid, L-glutamic acid, α-oxoglutaric acid, glycolic acid,hippuric acid, hydrobromic acid, hydrochloric acid, hydroiodic acid,(+)-L-lactic acid, (±)-DL-lactic acid, lactobionic acid, lauric acid,maleic acid, (−)-L-malic acid, malonic acid, (±)-DL-mandelic acid,methanesulfonic acid, naphthalene-2-sulfonic acid,naphthalene-1,5-disulfonic acid, 1-hydroxy-2-naphthoic acid, nicotinicacid, nitric acid, oleic acid, orotic acid, oxalic acid, palmitic acid,pamoic acid, perchloric acid, phosphoric acid, L-pyroglutamic acid,saccharic acid, salicylic acid, 4-amino-salicylic acid, sebacic acid,stearic acid, succinic acid, sulfuric acid, tannic acid, (+)-L-tartaricacid, thiocyanic acid, p-toluenesulfonic acid, undecylenic acid, andvaleric acid.

Suitable bases for use in the preparation of pharmaceutically acceptablesalts, include, but are not limited to, inorganic bases, such asmagnesium hydroxide, calcium hydroxide, potassium hydroxide, zinchydroxide, or sodium hydroxide; and organic bases, such as primary,secondary, tertiary, and quaternary, aliphatic and aromatic amines,including, but not limited to, L-arginine, benethamine, benzathine,choline, deanol, diethanolamine, diethylamine, dimethylamine,dipropylamine, diisopropylamine, 2-(diethylamino)-ethanol, ethanolamine,ethylamine, ethylenediamine, isopropylamine, N-methyl-glucamine,hydrabamine, 1H-imidazole, L-lysine, morpholine,4-(2-hydroxyethyl)-morpholine, methylamine, piperidine, piperazine,propylamine, pyrrolidine, 1-(2-hydroxyethyl)-pyrrolidine, pyridine,quinuclidine, quinoline, isoquinoline, secondary amines,triethanolamine, trimethylamine, triethylamine, N-methyl-D-glucamine,2-amino-2-(hydroxymethyl)-1,3-propanediol, and tromethamine.

5.3. Methods of Synthesis

The compounds provided herein can be prepared, isolated, or obtained byany methods known to one of skill in the art, and the following examplesare only representative and do not exclude other related procedures.

In one embodiment, for example, a compound of Formula I is prepared, asshown in Scheme 1 to form compound I.

5.4. Pharmaceutical Compositions

In one embodiment, provided herein is a pharmaceutical compositioncomprising the compound provided herein, including a compound of FormulaI, or an enantiomer, a mixture of enantiomers, a mixture of two or morediastereomers, or an isotopic variant thereof; or a pharmaceuticallyacceptable salt, solvate, or hydrate thereof; and a pharmaceuticallyacceptable excipient.

In one embodiment, the pharmaceutical composition provided herein isformulated in a dosage form for oral administration, which comprises acompound provided herein, and one or more pharmaceutically acceptableexcipients. In certain embodiments, the pharmaceutical composition fororal administration is formulated in tablet, capsule, powder, or liquidform.

In another embodiment, the pharmaceutical composition provided herein isformulated in a dosage form for parenteral administration, whichcomprises a compound provided herein, and one or more pharmaceuticallyacceptable excipients.

In yet another embodiment, the pharmaceutical composition providedherein is formulated in a dosage form for topical administration, whichcomprises a compound provided herein, and one or more pharmaceuticallyacceptable excipients.

In certain embodiments, the pharmaceutical composition provided hereinis formulated as modified release dosage forms, including delayed-,extended-, prolonged-, sustained-, pulsatile-, controlled-,accelerated-, fast-, targeted-, and programmed-release, and gastricretention dosage forms. These dosage forms can be prepared according toconventional methods and techniques known to those skilled in the art(see, Remington: The Science and Practice of Pharmacy, supra;Modified-Release Drug Delivery Technology, 2nd Edition, Rathbone et al.,Eds., Marcel Dekker, Inc.: New York, N.Y., 2008).

In certain embodiments, the pharmaceutical composition provided hereinis provided in a unit-dosage form or multiple-dosage form. A unit-dosageform, as used herein, refers to physically discrete a unit suitable foradministration to a human and animal subject, and packaged individuallyas is known in the art. Each unit-dose contains a predetermined quantityof an active ingredient(s) sufficient to produce the desired therapeuticeffect, in association with the required pharmaceutical carriers orexcipients. Examples of a unit-dosage form include an ampoule, syringe,and individually packaged tablet and capsule. A unit-dosage form may beadministered in fractions or multiples thereof. A multiple-dosage formis a plurality of identical unit-dosage forms packaged in a singlecontainer to be administered in segregated unit-dosage form. Examples ofa multiple-dosage form include a vial, bottle of tablets or capsules, orbottle of pints or gallons.

The pharmaceutical compositions provided herein can be administered atonce, or multiple times at intervals of time. It is understood that theprecise dosage and duration of treatment may vary with the age, weight,and condition of the patient being treated, and may be determinedempirically using known testing protocols or by extrapolation from invivo or in vitro test or diagnostic data. It is further understood thatfor any particular individual, specific dosage regimens should beadjusted over time according to the individual need and the professionaljudgment of the person administering or supervising the administrationof the formulations.

In certain embodiments, the pharmaceutical compositions provided hereinfurther comprise one or more therapeutic agents (e.g., chemotherapeuticagents) as defined herein.

5.4.1. Oral Administration

The pharmaceutical compositions provided herein for oral administrationcan be provided in solid, semisolid, or liquid dosage forms for oraladministration. As used herein, oral administration also includesbuccal, lingual, and sublingual administration. Suitable oral dosageforms include, but are not limited to, tablets, fastmelts, chewabletablets, capsules, pills, strips, troches, lozenges, pastilles, cachets,pellets, medicated chewing gum, bulk powders, effervescent ornon-effervescent powders or granules, oral mists, solutions, emulsions,suspensions, wafers, sprinkles, elixirs, and syrups. In addition to theactive ingredient(s), the pharmaceutical compositions can contain one ormore pharmaceutically acceptable carriers or excipients, including, butnot limited to, binders, fillers, diluents, disintegrants, wettingagents, lubricants, glidants, coloring agents, dye-migration inhibitors,sweetening agents, flavoring agents, emulsifying agents, suspending anddispersing agents, preservatives, solvents, non-aqueous liquids, organicacids, and sources of carbon dioxide.

Binders or granulators impart cohesiveness to a tablet to ensure thetablet remaining intact after compression. Suitable binders orgranulators include, but are not limited to, starches, such as cornstarch, potato starch, and pre-gelatinized starch (e.g., STARCH 1500);gelatin; sugars, such as sucrose, glucose, dextrose, molasses, andlactose; natural and synthetic gums, such as acacia, alginic acid,alginates, extract of Irish moss, panwar gum, ghatti gum, mucilage ofisabgol husks, carboxymethylcellulose, methylcellulose,polyvinylpyrrolidone (PVP), Veegum, larch arabogalactan, powderedtragacanth, and guar gum; celluloses, such as ethyl cellulose, celluloseacetate, carboxymethyl cellulose calcium, sodium carboxymethylcellulose, methyl cellulose, hydroxyethylcellulose (HEC),hydroxypropylcellulose (HPC), hydroxypropyl methyl cellulose (HPMC);microcrystalline celluloses, such as AVICEL-PH-101, AVICEL-PH-103,AVICEL RC-581, AVICEL-PH-105 (FMC Corp., Marcus Hook, Pa.); and mixturesthereof. Suitable fillers include, but are not limited to, talc, calciumcarbonate, microcrystalline cellulose, powdered cellulose, dextrates,kaolin, mannitol, silicic acid, sorbitol, starch, pre-gelatinizedstarch, and mixtures thereof. The amount of a binder or filler in thepharmaceutical compositions provided herein varies upon the type offormulation, and is readily discernible to those of ordinary skill inthe art. The binder or filler may be present from about 50 to about 99%by weight in the pharmaceutical compositions provided herein.

Suitable diluents include, but are not limited to, dicalcium phosphate,calcium sulfate, lactose, sorbitol, sucrose, inositol, cellulose,kaolin, mannitol, sodium chloride, dry starch, and powdered sugar.Certain diluents, such as mannitol, lactose, sorbitol, sucrose, andinositol, when present in sufficient quantity, can impart properties tosome compressed tablets that permit disintegration in the mouth bychewing. Such compressed tablets can be used as chewable tablets. Theamount of a diluent in the pharmaceutical compositions provided hereinvaries upon the type of formulation, and is readily discernible to thoseof ordinary skill in the art.

Suitable disintegrants include, but are not limited to, agar; bentonite;celluloses, such as methylcellulose and carboxymethylcellulose; woodproducts; natural sponge; cation-exchange resins; alginic acid; gums,such as guar gum and Veegum HV; citrus pulp; cross-linked celluloses,such as croscarmellose; cross-linked polymers, such as crospovidone;cross-linked starches; calcium carbonate; microcrystalline cellulose,such as sodium starch glycolate; polacrilin potassium; starches, such ascorn starch, potato starch, tapioca starch, and pre-gelatinized starch;clays; aligns; and mixtures thereof. The amount of a disintegrant in thepharmaceutical compositions provided herein varies upon the type offormulation, and is readily discernible to those of ordinary skill inthe art. The amount of a disintegrant in the pharmaceutical compositionsprovided herein varies upon the type of formulation, and is readilydiscernible to those of ordinary skill in the art. The pharmaceuticalcompositions provided herein may contain from about 0.5 to about 15% orfrom about 1 to about 5% by weight of a disintegrant.

Suitable lubricants include, but are not limited to, calcium stearate;magnesium stearate; mineral oil; light mineral oil; glycerin; sorbitol;mannitol; glycols, such as glycerol behenate and polyethylene glycol(PEG); stearic acid; sodium lauryl sulfate; talc; hydrogenated vegetableoil, including peanut oil, cottonseed oil, sunflower oil, sesame oil,olive oil, corn oil, and soybean oil; zinc stearate; ethyl oleate; ethyllaureate; agar; starch; lycopodium; silica or silica gels, such asAEROSIL® 200 (W.R. Grace Co., Baltimore, Md.) and CAB-O-SIL® (Cabot Co.of Boston, Mass.); and mixtures thereof. The pharmaceutical compositionsprovided herein may contain about 0.1 to about 5% by weight of alubricant.

Suitable glidants include, but are not limited to, colloidal silicondioxide, CAB-O-SIL® (Cabot Co. of Boston, Mass.), and asbestos-freetalc. Suitable coloring agents include, but are not limited to, any ofthe approved, certified, water soluble FD&C dyes, and water insolubleFD&C dyes suspended on alumina hydrate, and color lakes and mixturesthereof. A color lake is the combination by adsorption of awater-soluble dye to a hydrous oxide of a heavy metal, resulting in aninsoluble form of the dye. Suitable flavoring agents include, but arenot limited to, natural flavors extracted from plants, such as fruits,and synthetic blends of compounds which produce a pleasant tastesensation, such as peppermint and methyl salicylate. Suitable sweeteningagents include, but are not limited to, sucrose, lactose, mannitol,syrups, glycerin, and artificial sweeteners, such as saccharin andaspartame. Suitable emulsifying agents include, but are not limited to,gelatin, acacia, tragacanth, bentonite, and surfactants, such aspolyoxyethylene sorbitan monooleate (TWEEN® 20), polyoxyethylenesorbitan monooleate 80 (TWEEN® 80), and triethanolamine oleate. Suitablesuspending and dispersing agents include, but are not limited to, sodiumcarboxymethylcellulose, pectin, tragacanth, Veegum, acacia, sodiumcarbomethylcellulose, hydroxypropyl methyl cellulose, andpolyvinylpyrrolid one. Suitable preservatives include, but are notlimited to, glycerin, methyl and propylparaben, benzoic acid, sodiumbenzoate and alcohol. Suitable wetting agents include, but are notlimited to, propylene glycol monostearate, sorbitan monooleate,diethylene glycol monolaurate, and polyoxyethylene lauryl ether.Suitable solvents include, but are not limited to, glycerin, sorbitol,ethyl alcohol, and syrup. Suitable non-aqueous liquids utilized inemulsions include, but are not limited to, mineral oil and cottonseedoil. Suitable organic acids include, but are not limited to, citric andtartaric acid. Suitable sources of carbon dioxide include, but are notlimited to, sodium bicarbonate and sodium carbonate.

It should be understood that many carriers and excipients may serveseveral functions, even within the same formulation.

The pharmaceutical compositions provided herein for oral administrationcan be provided as compressed tablets, tablet triturates, chewablelozenges, rapidly dissolving tablets, multiple compressed tablets, orenteric-coating tablets, sugar-coated, or film-coated tablets.Enteric-coated tablets are compressed tablets coated with substancesthat resist the action of stomach acid but dissolve or disintegrate inthe intestine, thus protecting the active ingredients from the acidicenvironment of the stomach. Enteric-coatings include, but are notlimited to, fatty acids, fats, phenyl salicylate, waxes, shellac,ammoniated shellac, and cellulose acetate phthalates. Sugar-coatedtablets are compressed tablets surrounded by a sugar coating, which maybe beneficial in covering up objectionable tastes or odors and inprotecting the tablets from oxidation. Film-coated tablets arecompressed tablets that are covered with a thin layer or film of awater-soluble material. Film coatings include, but are not limited to,hydroxyethylcellulose, sodium carboxymethylcellulose, polyethyleneglycol 4000, and cellulose acetate phthalate. Film coating imparts thesame general characteristics as sugar coating. Multiple compressedtablets are compressed tablets made by more than one compression cycle,including layered tablets, and press-coated or dry-coated tablets.

The tablet dosage forms can be prepared from the active ingredient inpowdered, crystalline, or granular forms, alone or in combination withone or more carriers or excipients described herein, including binders,disintegrants, controlled-release polymers, lubricants, diluents, and/orcolorants. Flavoring and sweetening agents are especially useful in theformation of chewable tablets and lozenges.

The pharmaceutical compositions provided herein for oral administrationcan be provided as soft or hard capsules, which can be made fromgelatin, methylcellulose, starch, or calcium alginate. The hard gelatincapsule, also known as the dry-filled capsule (DFC), consists of twosections, one slipping over the other, thus completely enclosing theactive ingredient. The soft elastic capsule (SEC) is a soft, globularshell, such as a gelatin shell, which is plasticized by the addition ofglycerin, sorbitol, or a similar polyol. The soft gelatin shells maycontain a preservative to prevent the growth of microorganisms. Suitablepreservatives are those as described herein, including methyl- andpropyl-parabens, and sorbic acid. The liquid, semisolid, and soliddosage forms provided herein may be encapsulated in a capsule. Suitableliquid and semisolid dosage forms include solutions and suspensions inpropylene carbonate, vegetable oils, or triglycerides. Capsulescontaining such solutions can be prepared as described in U.S. Pat. Nos.4,328,245; 4,409,239; and 4,410,545. The capsules may also be coated asknown by those of skill in the art in order to modify or sustaindissolution of the active ingredient.

The pharmaceutical compositions provided herein for oral administrationcan be provided in liquid and semisolid dosage forms, includingemulsions, solutions, suspensions, elixirs, and syrups. An emulsion is atwo-phase system, in which one liquid is dispersed in the form of smallglobules throughout another liquid, which can be oil-in-water orwater-in-oil. Emulsions may include a pharmaceutically acceptablenon-aqueous liquid or solvent, emulsifying agent, and preservative.Suspensions may include a pharmaceutically acceptable suspending agentand preservative. Aqueous alcoholic solutions may include apharmaceutically acceptable acetal, such as a di(lower alkyl) acetal ofa lower alkyl aldehyde, e.g., acetaldehyde diethyl acetal; and awater-miscible solvent having one or more hydroxyl groups, such aspropylene glycol and ethanol. Elixirs are clear, sweetened, andhydroalcoholic solutions. Syrups are concentrated aqueous solutions of asugar, for example, sucrose, and may also contain a preservative. For aliquid dosage form, for example, a solution in a polyethylene glycol maybe diluted with a sufficient quantity of a pharmaceutically acceptableliquid carrier, e.g., water, to be measured conveniently foradministration.

Other useful liquid and semisolid dosage forms include, but are notlimited to, those containing the active ingredient(s) provided herein,and a dialkylated mono- or poly-alkylene glycol, including,1,2-dimethoxymethane, diglyme, triglyme, tetraglyme, polyethyleneglycol-350-dimethyl ether, polyethylene glycol-550-dimethyl ether,polyethylene glycol-750-dimethyl ether, wherein 350, 550, and 750 referto the approximate average molecular weight of the polyethylene glycol.These formulations can further comprise one or more antioxidants, suchas butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA),propyl gallate, vitamin E, hydroquinone, hydroxycoumarins, ethanolamine,lecithin, cephalin, ascorbic acid, malic acid, sorbitol, phosphoricacid, bisulfite, sodium metabisulfite, thiodipropionic acid and itsesters, and dithiocarbamates.

The pharmaceutical compositions provided herein for oral administrationcan be also provided in the forms of liposomes, micelles, microspheres,or nanosystems. Micellar dosage forms can be prepared as described inU.S. Pat. No. 6,350,458.

The pharmaceutical compositions provided herein for oral administrationcan be provided as non-effervescent or effervescent, granules andpowders, to be reconstituted into a liquid dosage form. Pharmaceuticallyacceptable carriers and excipients used in the non-effervescent granulesor powders may include diluents, sweeteners, and wetting agents.Pharmaceutically acceptable carriers and excipients used in theeffervescent granules or powders may include organic acids and a sourceof carbon dioxide.

Coloring and flavoring agents can be used in all of the above dosageforms.

The pharmaceutical compositions provided herein for oral administrationcan be formulated as immediate or modified release dosage forms,including delayed-, sustained, pulsed-, controlled, targeted-, andprogrammed-release forms.

5.4.2. Parenteral Administration

The pharmaceutical compositions provided herein can be administeredparenterally by injection, infusion, or implantation, for local orsystemic administration. Parenteral administration, as used herein,include intravenous, intraarterial, intraperitoneal, intrathecal,intraventricular, intraurethral, intrasternal, intracranial,intramuscular, intrasynovial, intravesical, and subcutaneousadministration.

The pharmaceutical compositions provided herein for parenteraladministration can be formulated in any dosage forms that are suitablefor parenteral administration, including solutions, suspensions,emulsions, micelles, liposomes, microspheres, nanosystems, and solidforms suitable for solutions or suspensions in liquid prior toinjection. Such dosage forms can be prepared according to conventionalmethods known to those skilled in the art of pharmaceutical science(see, Remington: The Science and Practice of Pharmacy, supra).

The pharmaceutical compositions intended for parenteral administrationcan include one or more pharmaceutically acceptable carriers andexcipients, including, but not limited to, aqueous vehicles,water-miscible vehicles, non-aqueous vehicles, antimicrobial agents orpreservatives against the growth of microorganisms, stabilizers,solubility enhancers, isotonic agents, buffering agents, antioxidants,local anesthetics, suspending and dispersing agents, wetting oremulsifying agents, complexing agents, sequestering or chelating agents,cryoprotectants, lyoprotectants, thickening agents, pH adjusting agents,and inert gases.

Suitable aqueous vehicles include, but are not limited to, water,saline, physiological saline or phosphate buffered saline (PBS), sodiumchloride injection, Ringers injection, isotonic dextrose injection,sterile water injection, dextrose and lactated Ringers injection.Suitable non-aqueous vehicles include, but are not limited to, fixedoils of vegetable origin, castor oil, corn oil, cottonseed oil, oliveoil, peanut oil, peppermint oil, safflower oil, sesame oil, soybean oil,hydrogenated vegetable oils, hydrogenated soybean oil, and medium-chaintriglycerides of coconut oil, and palm seed oil. Suitable water-misciblevehicles include, but are not limited to, ethanol, 1,3-butanediol,liquid polyethylene glycol (e.g., polyethylene glycol 300 andpolyethylene glycol 400), propylene glycol, glycerin,N-methyl-2-pyrrolidone, N,N-dimethylacetamide, and dimethyl sulfoxide.

Suitable antimicrobial agents or preservatives include, but are notlimited to, phenols, cresols, mercurials, benzyl alcohol, chlorobutanol,methyl and propyl p-hydroxybenzoates, thimerosal, benzalkonium chloride(e.g., benzethonium chloride), methyl- and propyl-parabens, and sorbicacid. Suitable isotonic agents include, but are not limited to, sodiumchloride, glycerin, and dextrose. Suitable buffering agents include, butare not limited to, phosphate and citrate. Suitable antioxidants arethose as described herein, including bisulfite and sodium metabisulfite.Suitable local anesthetics include, but are not limited to, procainehydrochloride. Suitable suspending and dispersing agents are those asdescribed herein, including sodium carboxymethylcelluose, hydroxypropylmethylcellulose, and polyvinylpyrrolidone. Suitable emulsifying agentsare those described herein, including polyoxyethylene sorbitanmonolaurate, polyoxyethylene sorbitan monooleate 80, and triethanolamineoleate. Suitable sequestering or chelating agents include, but are notlimited to EDTA. Suitable pH adjusting agents include, but are notlimited to, sodium hydroxide, hydrochloric acid, citric acid, and lacticacid. Suitable complexing agents include, but are not limited to,cyclodextrins, including α-cyclodextrin, β-cyclodextrin,hydroxypropyl-β-cyclodextrin, sulfobutylether-β-cyclodextrin, andsulfobutylether 7-β-cyclodextrin (CAPTISOL®, CyDex, Lenexa, Kans.).

When the pharmaceutical compositions provided herein are formulated formultiple dosage administration, the multiple dosage parenteralformulations must contain an antimicrobial agent at bacteriostatic orfungistatic concentrations. All parenteral formulations must be sterile,as known and practiced in the art.

In one embodiment, the pharmaceutical compositions for parenteraladministration are provided as ready-to-use sterile solutions. Inanother embodiment, the pharmaceutical compositions are provided assterile dry soluble products, including lyophilized powders andhypodermic tablets, to be reconstituted with a vehicle prior to use. Inyet another embodiment, the pharmaceutical compositions are provided asready-to-use sterile suspensions. In yet another embodiment, thepharmaceutical compositions are provided as sterile dry insolubleproducts to be reconstituted with a vehicle prior to use. In stillanother embodiment, the pharmaceutical compositions are provided asready-to-use sterile emulsions.

The pharmaceutical compositions provided herein for parenteraladministration can be formulated as immediate or modified release dosageforms, including delayed-, sustained, pulsed-, controlled, targeted-,and programmed-release forms.

The pharmaceutical compositions provided herein for parenteraladministration can be formulated as a suspension, solid, semi-solid, orthixotropic liquid, for administration as an implanted depot. In oneembodiment, the pharmaceutical compositions provided herein aredispersed in a solid inner matrix, which is surrounded by an outerpolymeric membrane that is insoluble in body fluids but allows theactive ingredient in the pharmaceutical compositions diffuse through.

Suitable inner matrixes include, but are not limited to,polymethylmethacrylate, polybutyl-methacrylate, plasticized orunplasticized polyvinylchloride, plasticized nylon, plasticizedpolyethylene terephthalate, natural rubber, polyisoprene,polyisobutylene, polybutadiene, polyethylene, ethylene-vinyl acetatecopolymers, silicone rubbers, polydimethylsiloxanes, silicone carbonatecopolymers, hydrophilic polymers, such as hydrogels of esters of acrylicand methacrylic acid, collagen, cross-linked polyvinyl alcohol, andcross-linked partially hydrolyzed polyvinyl acetate.

Suitable outer polymeric membranes include but are not limited to,polyethylene, polypropylene, ethylene/propylene copolymers,ethylene/ethyl acrylate copolymers, ethylene/vinyl acetate copolymers,silicone rubbers, polydimethyl siloxanes, neoprene rubber, chlorinatedpolyethylene, polyvinylchloride, vinyl chloride copolymers with vinylacetate, vinylidene chloride, ethylene and propylene, ionomerpolyethylene terephthalate, butyl rubber epichlorohydrin rubbers,ethylene/vinyl alcohol copolymer, ethylene/vinyl acetate/vinyl alcoholterpolymer, and ethylene/vinyloxyethanol copolymer.

5.4.3. Topical Administration

The pharmaceutical compositions provided herein can be administeredtopically to the skin, orifices, or mucosa. The topical administration,as used herein, includes (intra)dermal, conjunctival, intracorneal,intraocular, ophthalmic, auricular, transdermal, nasal, vaginal,urethral, respiratory, and rectal administration.

The pharmaceutical compositions provided herein can be formulated in anydosage forms that are suitable for topical administration for local orsystemic effect, including emulsions, solutions, suspensions, creams,gels, hydrogels, ointments, dusting powders, dressings, elixirs,lotions, suspensions, tinctures, pastes, foams, films, aerosols,irrigations, sprays, suppositories, bandages, and dermal patches. Thetopical formulation of the pharmaceutical compositions provided hereincan also comprise liposomes, micelles, microspheres, nanosystems, andmixtures thereof.

Pharmaceutically acceptable carriers and excipients suitable for use inthe topical formulations provided herein include, but are not limitedto, aqueous vehicles, water-miscible vehicles, non-aqueous vehicles,antimicrobial agents or preservatives against the growth ofmicroorganisms, stabilizers, solubility enhancers, isotonic agents,buffering agents, antioxidants, local anesthetics, suspending anddispersing agents, wetting or emulsifying agents, complexing agents,sequestering or chelating agents, penetration enhancers,cryoprotectants, lyoprotectants, thickening agents, and inert gases.

The pharmaceutical compositions can also be administered topically byelectroporation, iontophoresis, phonophoresis, sonophoresis, ormicroneedle or needle-free injection, such as POWDERJECT™ (Chiron Corp.,Emeryville, Calif.), and BIOJECT™ (Bioject Medical Technologies Inc.,Tualatin, Oreg.).

The pharmaceutical compositions provided herein can be provided in theforms of ointments, creams, and gels. Suitable ointment vehicles includeoleaginous or hydrocarbon vehicles, including lard, benzoinated lard,olive oil, cottonseed oil, and other oils, white petrolatum;emulsifiable or absorption vehicles, such as hydrophilic petrolatum,hydroxystearin sulfate, and anhydrous lanolin; water-removable vehicles,such as hydrophilic ointment; water-soluble ointment vehicles, includingpolyethylene glycols of varying molecular weight; emulsion vehicles,either water-in-oil (W/O) emulsions or oil-in-water (O/W) emulsions,including cetyl alcohol, glyceryl monostearate, lanolin, and stearicacid (see, Remington: The Science and Practice of Pharmacy, supra).These vehicles are emollient but generally require addition ofantioxidants and preservatives.

Suitable cream base can be oil-in-water or water-in-oil. Suitable creamvehicles may be water-washable, and contain an oil phase, an emulsifier,and an aqueous phase. The oil phase is also called the “internal” phase,which is generally comprised of petrolatum and a fatty alcohol such ascetyl or stearyl alcohol. The aqueous phase usually, although notnecessarily, exceeds the oil phase in volume, and generally contains ahumectant. The emulsifier in a cream formulation may be a nonionic,anionic, cationic, or amphoteric surfactant.

Gels are semisolid, suspension-type systems. Single-phase gels containorganic macromolecules distributed substantially uniformly throughoutthe liquid carrier. Suitable gelling agents include, but are not limitedto, crosslinked acrylic acid polymers, such as carbomers,carboxypolyalkylenes, and CARBOPOL®; hydrophilic polymers, such aspolyethylene oxides, polyoxyethylene-polyoxypropylene copolymers, andpolyvinylalcohol; cellulosic polymers, such as hydroxypropyl cellulose,hydroxyethyl cellulose, hydroxypropyl methylcellulose, hydroxypropylmethylcellulose phthalate, and methylcellulose; gums, such as tragacanthand xanthan gum; sodium alginate; and gelatin. In order to prepare auniform gel, dispersing agents such as alcohol or glycerin can be added,or the gelling agent can be dispersed by trituration, mechanical mixing,and/or stirring.

The pharmaceutical compositions provided herein can be administeredrectally, urethrally, vaginally, or perivaginally in the forms ofsuppositories, pessaries, bougies, poultices or cataplasm, pastes,powders, dressings, creams, plasters, contraceptives, ointments,solutions, emulsions, suspensions, tampons, gels, foams, sprays, orenemas. These dosage forms can be manufactured using conventionalprocesses as described in Remington: The Science and Practice ofPharmacy, supra.

Rectal, urethral, and vaginal suppositories are solid bodies forinsertion into body orifices, which are solid at ordinary temperaturesbut melt or soften at body temperature to release the activeingredient(s) inside the orifices. Pharmaceutically acceptable carriersutilized in rectal and vaginal suppositories include bases or vehicles,such as stiffening agents, which produce a melting point in theproximity of body temperature, when formulated with the pharmaceuticalcompositions provided herein; and antioxidants as described herein,including bisulfite and sodium metabisulfite. Suitable vehicles include,but are not limited to, cocoa butter (theobroma oil), glycerin-gelatin,carbowax (polyoxyethylene glycol), spermaceti, paraffin, white andyellow wax, and appropriate mixtures of mono-, di- and triglycerides offatty acids, and hydrogels, such as polyvinyl alcohol, hydroxyethylmethacrylate, and polyacrylic acid. Combinations of the various vehiclescan also be used. Rectal and vaginal suppositories may be prepared bycompressing or molding. The typical weight of a rectal and vaginalsuppository is about 2 to about 3 g.

The pharmaceutical compositions provided herein can be administeredophthalmically in the forms of solutions, suspensions, ointments,emulsions, gel-forming solutions, powders for solutions, gels, ocularinserts, and implants.

The pharmaceutical compositions provided herein can be administeredintranasally or by inhalation to the respiratory tract. Thepharmaceutical compositions can be provided in the form of an aerosol orsolution for delivery using a pressurized container, pump, spray,atomizer, such as an atomizer using electrohydrodynamics to produce afine mist, or nebulizer, alone or in combination with a suitablepropellant, such as 1,1,1,2-tetrafluoroethane or1,1,1,2,3,3,3-heptafluoropropane. The pharmaceutical compositions canalso be provided as a dry powder for insufflation, alone or incombination with an inert carrier such as lactose or phospholipids; andnasal drops. For intranasal use, the powder can comprise a bioadhesiveagent, including chitosan or cyclodextrin.

Solutions or suspensions for use in a pressurized container, pump,spray, atomizer, or nebulizer can be formulated to contain ethanol,aqueous ethanol, or a suitable alternative agent for dispersing,solubilizing, or extending release of the active ingredient providedherein; a propellant as solvent; and/or a surfactant, such as sorbitantrioleate, oleic acid, or an oligolactic acid.

The pharmaceutical compositions provided herein can be micronized to asize suitable for delivery by inhalation, such as about 50 micrometersor less, or about 10 micrometers or less. Particles of such sizes can beprepared using a comminuting method known to those skilled in the art,such as spiral jet milling, fluid bed jet milling, supercritical fluidprocessing to form nanoparticles, high pressure homogenization, or spraydrying.

Capsules, blisters, and cartridges for use in an inhaler or insufflatorcan be formulated to contain a powder mix of the pharmaceuticalcompositions provided herein; a suitable powder base, such as lactose orstarch; and a performance modifier, such as l-leucine, mannitol, ormagnesium stearate. The lactose may be anhydrous or in the form of themonohydrate. Other suitable excipients or carriers include, but are notlimited to, dextran, glucose, maltose, sorbitol, xylitol, fructose,sucrose, and trehalose. The pharmaceutical compositions provided hereinfor inhaled/intranasal administration can further comprise a suitableflavor, such as menthol and levomenthol; and/or sweeteners, such assaccharin and saccharin sodium.

The pharmaceutical compositions provided herein for topicaladministration can be formulated to be immediate release or modifiedrelease, including delayed-, sustained-, pulsed-, controlled-, targeted,and programmed release.

5.4.4. Modified Release

The pharmaceutical compositions provided herein can be formulated as amodified release dosage form. As used herein, the term “modifiedrelease” refers to a dosage form in which the rate or place of releaseof the active ingredient(s) is different from that of an immediatedosage form when administered by the same route. Modified release dosageforms include, but are not limited to, delayed-, extended-, prolonged-,sustained-, pulsatile-, controlled-, accelerated- and fast-, targeted-,programmed-release, and gastric retention dosage forms. Thepharmaceutical compositions in modified release dosage forms can beprepared using a variety of modified release devices and methods knownto those skilled in the art, including, but not limited to, matrixcontrolled release devices, osmotic controlled release devices,multiparticulate controlled release devices, ion-exchange resins,enteric coatings, multilayered coatings, microspheres, liposomes, andcombinations thereof. The release rate of the active ingredient(s) canalso be modified by varying the particle sizes and polymorphorism of theactive ingredient(s).

Examples of modified release include, but are not limited to, thosedescribed in U.S. Pat. Nos. 3,845,770; 3,916,899; 3,536,809; 3,598,123;4,008,719; 5,674,533; 5,059,595; 5,591,767; 5,120,548; 5,073,543;5,639,476; 5,354,556; 5,639,480; 5,733,566; 5,739,108; 5,891,474;5,922,356; 5,972,891; 5,980,945; 5,993,855; 6,045,830; 6,087,324;6,113,943; 6,197,350; 6,248,363; 6,264,970; 6,267,981; 6,376,461;6,419,961; 6,589,548; 6,613,358; and 6,699,500.

5.4.4.1. Matrix Controlled Release Devices

The pharmaceutical compositions provided herein in a modified releasedosage form can be fabricated using a matrix controlled release deviceknown to those skilled in the art (see, Takada et al. in “Encyclopediaof Controlled Drug Delivery,” Vol. 2, Mathiowitz Ed., Wiley, 1999).

In certain embodiments, the pharmaceutical compositions provided hereinin a modified release dosage form is formulated using an erodible matrixdevice, which is water-swellable, erodible, or soluble polymers,including, but not limited to, synthetic polymers, and naturallyoccurring polymers and derivatives, such as polysaccharides andproteins.

Materials useful in forming an erodible matrix include, but are notlimited to, chitin, chitosan, dextran, and pullulan; gum agar, gumarabic, gum karaya, locust bean gum, gum tragacanth, carrageenans, gumghatti, guar gum, xanthan gum, and scleroglucan; starches, such asdextrin and maltodextrin; hydrophilic colloids, such as pectin;phosphatides, such as lecithin; alginates; propylene glycol alginate;gelatin; collagen; cellulosics, such as ethyl cellulose (EC),methylethyl cellulose (MEC), carboxymethyl cellulose (CMC), CMEC,hydroxyethyl cellulose (HEC), hydroxypropyl cellulose (HPC), celluloseacetate (CA), cellulose propionate (CP), cellulose butyrate (CB),cellulose acetate butyrate (CAB), CAP, CAT, hydroxypropyl methylcellulose (HPMC), HPMCP, HPMCAS, hydroxypropyl methyl cellulose acetatetrimellitate (HPMCAT), and ethyl hydroxyethyl cellulose (EHEC);polyvinyl pyrrolidone; polyvinyl alcohol; polyvinyl acetate; glycerolfatty acid esters; polyacrylamide; polyacrylic acid; copolymers ofethacrylic acid or methacrylic acid (EUDRAGIT®, Rohm America, Inc.,Piscataway, N.J.); poly(2-hydroxyethyl-methacrylate); polylactides;copolymers of L-glutamic acid and ethyl-L-glutamate; degradable lacticacid-glycolic acid copolymers; poly-D-(−)-3-hydroxybutyric acid; andother acrylic acid derivatives, such as homopolymers and copolymers ofbutylmethacrylate, methyl methacrylate, ethyl methacrylate,ethylacrylate, (2-dimethylaminoethyl)methacrylate, and(trimethylaminoethyl)methacrylate chloride.

In certain embodiments, the pharmaceutical compositions provided hereinare formulated with a non-erodible matrix device. The activeingredient(s) is dissolved or dispersed in an inert matrix and isreleased primarily by diffusion through the inert matrix onceadministered. Materials suitable for use as a non-erodible matrix deviceinclude, but are not limited to, insoluble plastics, such aspolyethylene, polypropylene, polyisoprene, polyisobutylene,polybutadiene, polymethylmethacrylate, polybutylmethacrylate,chlorinated polyethylene, polyvinylchloride, methyl acrylate-methylmethacrylate copolymers, ethylene-vinyl acetate copolymers,ethylene/propylene copolymers, ethylene/ethyl acrylate copolymers, vinylchloride copolymers with vinyl acetate, vinylidene chloride, ethyleneand propylene, ionomer polyethylene terephthalate, butyl rubbers,epichlorohydrin rubbers, ethylene/vinyl alcohol copolymer,ethylene/vinyl acetate/vinyl alcohol terpolymer,ethylene/vinyloxyethanol copolymer, polyvinyl chloride, plasticizednylon, plasticized polyethylene terephthalate, natural rubber, siliconerubbers, polydimethylsiloxanes, and silicone carbonate copolymers;hydrophilic polymers, such as ethyl cellulose, cellulose acetate,crospovidone, and cross-linked partially hydrolyzed polyvinyl acetate;and fatty compounds, such as carnauba wax, microcrystalline wax, andtriglycerides.

In a matrix controlled release system, the desired release kinetics canbe controlled, for example, via the polymer type employed, the polymerviscosity, the particle sizes of the polymer and/or the activeingredient(s), the ratio of the active ingredient(s) versus the polymer,and other excipients or carriers in the compositions.

The pharmaceutical compositions provided herein in a modified releasedosage form can be prepared by methods known to those skilled in theart, including direct compression, dry or wet granulation followed bycompression, and melt-granulation followed by compression.

5.4.4.2. Osmotic Controlled Release Devices

The pharmaceutical compositions provided herein in a modified releasedosage form can be fabricated using an osmotic controlled releasedevice, including, but not limited to, one-chamber system, two-chambersystem, asymmetric membrane technology (AMT), and extruding core system(ECS). In general, such devices have at least two components: (a) a corewhich contains an active ingredient; and (b) a semipermeable membranewith at least one delivery port, which encapsulates the core. Thesemipermeable membrane controls the influx of water to the core from anaqueous environment of use so as to cause drug release by extrusionthrough the delivery port(s).

In addition to the active ingredient(s), the core of the osmotic deviceoptionally includes an osmotic agent, which creates a driving force fortransport of water from the environment of use into the core of thedevice. One class of osmotic agents is water-swellable hydrophilicpolymers, which are also referred to as “osmopolymers” and “hydrogels.”Suitable water-swellable hydrophilic polymers as osmotic agents include,but are not limited to, hydrophilic vinyl and acrylic polymers,polysaccharides such as calcium alginate, polyethylene oxide (PEO),polyethylene glycol (PEG), polypropylene glycol (PPG),poly(2-hydroxyethyl methacrylate), poly(acrylic) acid, poly(methacrylic)acid, polyvinylpyrrolidone (PVP), crosslinked PVP, polyvinyl alcohol(PVA), PVA/PVP copolymers, PVA/PVP copolymers with hydrophobic monomerssuch as methyl methacrylate and vinyl acetate, hydrophilic polyurethanescontaining large PEO blocks, sodium croscarmellose, carrageenan,hydroxyethyl cellulose (HEC), hydroxypropyl cellulose (HPC),hydroxypropyl methyl cellulose (HPMC), carboxymethyl cellulose (CMC) andcarboxyethyl, cellulose (CEC), sodium alginate, polycarbophil, gelatin,xanthan gum, and sodium starch glycolate.

The other class of osmotic agents is osmogens, which are capable ofimbibing water to affect an osmotic pressure gradient across the barrierof the surrounding coating. Suitable osmogens include, but are notlimited to, inorganic salts, such as magnesium sulfate, magnesiumchloride, calcium chloride, sodium chloride, lithium chloride, potassiumsulfate, potassium phosphates, sodium carbonate, sodium sulfite, lithiumsulfate, potassium chloride, and sodium sulfate; sugars, such asdextrose, fructose, glucose, inositol, lactose, maltose, mannitol,raffinose, sorbitol, sucrose, trehalose, and xylitol; organic acids,such as ascorbic acid, benzoic acid, fumaric acid, citric acid, maleicacid, sebacic acid, sorbic acid, adipic acid, edetic acid, glutamicacid, p-toluenesulfonic acid, succinic acid, and tartaric acid; urea;and mixtures thereof.

Osmotic agents of different dissolution rates can be employed toinfluence how rapidly the active ingredient(s) is initially deliveredfrom the dosage form. For example, amorphous sugars, such as MANNOGEM™EZ (SPI Pharma, Lewes, Del.) can be used to provide faster deliveryduring the first couple of hours to promptly produce the desiredtherapeutic effect, and gradually and continually release of theremaining amount to maintain the desired level of therapeutic orprophylactic effect over an extended period of time. In this case, theactive ingredient(s) is released at such a rate to replace the amount ofthe active ingredient metabolized and excreted.

The core can also include a wide variety of other excipients andcarriers as described herein to enhance the performance of the dosageform or to promote stability or processing.

Materials useful in forming the semipermeable membrane include variousgrades of acrylics, vinyls, ethers, polyamides, polyesters, andcellulosic derivatives that are water-permeable and water-insoluble atphysiologically relevant pHs, or are susceptible to being renderedwater-insoluble by chemical alteration, such as crosslinking. Examplesof suitable polymers useful in forming the coating, include plasticized,unplasticized, and reinforced cellulose acetate (CA), cellulosediacetate, cellulose triacetate, CA propionate, cellulose nitrate,cellulose acetate butyrate (CAB), CA ethyl carbamate, CAP, CA methylcarbamate, CA succinate, cellulose acetate trimellitate (CAT), CAdimethylaminoacetate, CA ethyl carbonate, CA chloroacetate, CA ethyloxalate, CA methyl sulfonate, CA butyl sulfonate, CA p-toluenesulfonate, agar acetate, amylose triacetate, beta glucan acetate, betaglucan triacetate, acetaldehyde dimethyl acetate, triacetate of locustbean gum, hydroxylated ethylene-vinylacetate, EC, PEG, PPG, PEG/PPGcopolymers, PVP, HEC, HPC, CMC, CMEC, HPMC, HPMCP, HPMCAS, HPMCAT,poly(acrylic) acids and esters and poly-(methacrylic) acids and estersand copolymers thereof, starch, dextran, dextrin, chitosan, collagen,gelatin, polyalkenes, polyethers, polysulfones, polyethersulfones,polystyrenes, polyvinyl halides, polyvinyl esters and ethers, naturalwaxes, and synthetic waxes.

Semipermeable membrane can also be a hydrophobic microporous membrane,wherein the pores are substantially filled with a gas and are not wettedby the aqueous medium but are permeable to water vapor, as disclosed inU.S. Pat. No. 5,798,119. Such hydrophobic but water-vapor permeablemembrane are typically composed of hydrophobic polymers such aspolyalkenes, polyethylene, polypropylene, polytetrafluoroethylene,polyacrylic acid derivatives, polyethers, polysulfones,polyethersulfones, polystyrenes, polyvinyl halides, polyvinylidenefluoride, polyvinyl esters and ethers, natural waxes, and syntheticwaxes.

The delivery port(s) on the semipermeable membrane can be formedpost-coating by mechanical or laser drilling. Delivery port(s) can alsobe formed in situ by erosion of a plug of water-soluble material or byrupture of a thinner portion of the membrane over an indentation in thecore. In addition, delivery ports can be formed during coating process,as in the case of asymmetric membrane coatings of the type disclosed inU.S. Pat. Nos. 5,612,059 and 5,698,220.

The total amount of the active ingredient(s) released and the releaserate can substantially by modulated via the thickness and porosity ofthe semipermeable membrane, the composition of the core, and the number,size, and position of the delivery ports.

The pharmaceutical compositions in an osmotic controlled-release dosageform can further comprise additional conventional excipients or carriersas described herein to promote performance or processing of theformulation.

The osmotic controlled-release dosage forms can be prepared according toconventional methods and techniques known to those skilled in the art(see, Remington: The Science and Practice of Pharmacy, supra; Santus andBaker, J. Controlled Release 1995, 35, 1-21; Verma et al., DrugDevelopment and Industrial Pharmacy 2000, 26, 695-708; Verma et al., J.Controlled Release 2002, 79, 7-27).

In certain embodiments, the pharmaceutical compositions provided hereinare formulated as AMT controlled-release dosage form, which comprises anasymmetric osmotic membrane that coats a core comprising the activeingredient(s) and other pharmaceutically acceptable excipients orcarriers. See, U.S. Pat. No. 5,612,059 and WO 2002/17918. The AMTcontrolled-release dosage forms can be prepared according toconventional methods and techniques known to those skilled in the art,including direct compression, dry granulation, wet granulation, and adip-coating method.

In certain embodiments, the pharmaceutical compositions provided hereinare formulated as ESC controlled-release dosage form, which comprises anosmotic membrane that coats a core comprising the active ingredient(s),a hydroxylethyl cellulose, and other pharmaceutically acceptableexcipients or carriers.

5.4.4.3. Multiparticulate Controlled Release Devices

The pharmaceutical compositions provided herein in a modified releasedosage form can be fabricated as a multiparticulate controlled releasedevice, which comprises a multiplicity of particles, granules, orpellets, ranging from about 10 μm to about 3 mm, about 50 μm to about2.5 mm, or from about 100 μm to about 1 mm in diameter. Suchmultiparticulates can be made by the processes known to those skilled inthe art, including wet- and dry-granulation, extrusion/spheronization,roller-compaction, melt-congealing, and by spray-coating seed cores.See, for example, Multiparticulate Oral Drug Delivery; Marcel Dekker:1994; and Pharmaceutical Pelletization Technology; Marcel Dekker: 1989.

Other excipients or carriers as described herein can be blended with thepharmaceutical compositions to aid in processing and forming themultiparticulates. The resulting particles can themselves constitute themultiparticulate device or can be coated by various film-formingmaterials, such as enteric polymers, water-swellable, and water-solublepolymers. The multiparticulates can be further processed as a capsule ora tablet.

5.4.4.4. Targeted Delivery

The pharmaceutical compositions provided herein can also be formulatedto be targeted to a particular tissue, receptor, or other area of thebody of the subject to be treated, including liposome-, resealederythrocyte-, and antibody-based delivery systems. Examples include, butare not limited to, those disclosed in U.S. Pat. Nos. 6,316,652;6,274,552; 6,271,359; 6,253,872; 6,139,865; 6,131,570; 6,120,751;6,071,495; 6,060,082; 6,048,736; 6,039,975; 6,004,534; 5,985,307;5,972,366; 5,900,252; 5,840,674; 5,759,542; and 5,709,874.

5.5. Articles of Manufacture

In one embodiment, the compounds and the pharmaceutical compositionsprovided herein are provided as an article of manufacture usingpackaging materials well known to those of skill in the art. See, e.g.,U.S. Pat. Nos. 5,323,907; 5,052,558; and 5,033,252. Examples ofpharmaceutical packaging materials include, but are not limited to,blister packs, bottles, tubes, inhalers, pumps, bags, vials, containers,syringes, and any packaging material suitable for a selected formulationand intended mode of administration and treatment.

In another embodiment, provided herein are kits which, when used by themedical practitioner, can simplify the administration of appropriateamounts of active ingredients to a subject. In certain embodiments, thekit provided herein includes a container and a dosage form of a compoundprovided herein, e.g., a compound of Formula I, or an enantiomer, amixture of enantiomers, a mixture of two or more diastereomers, or anisotopic variant thereof; or a pharmaceutically acceptable salt,solvate, or hydrate thereof.

In certain embodiments, the kit includes a container comprising a dosageform of the compound provided herein, e.g., a compound of Formula I, oran enantiomer, a mixture of enantiomers, a mixture of two or morediastereomers, or an isotopic variant thereof; or a pharmaceuticallyacceptable salt, solvate, hydrate, or prodrug thereof, in a containercomprising one or more other therapeutic agent(s) described herein.

Kits provided herein can further include devices that are used toadminister the active ingredients. Examples of such devices include, butare not limited to, syringes, needle-less injectors drip bags, patches,and inhalers. The kits provided herein can also include condoms foradministration of the active ingredients.

Kits provided herein can further include pharmaceutically acceptablevehicles that can be used to administer one or more active ingredients.For example, if an active ingredient is provided in a solid form thatmust be reconstituted for parenteral administration, the kit cancomprise a sealed container of a suitable vehicle in which the activeingredient can be dissolved to form a particulate-free sterile solutionthat is suitable for parenteral administration. Examples ofpharmaceutically acceptable vehicles include, but are not limited to:aqueous vehicles, including, but not limited to, Water for InjectionUSP, Sodium Chloride Injection, Ringer's Injection, Dextrose Injection,Dextrose and Sodium Chloride Injection, and Lactated Ringer's Injection;water-miscible vehicles, including, but not limited to, ethyl alcohol,polyethylene glycol, and polypropylene glycol; and non-aqueous vehicles,including, but not limited to, corn oil, cottonseed oil, peanut oil,sesame oil, ethyl oleate, isopropyl myristate, and benzyl benzoate.

5.6. Methods of Treatment

In one embodiment, provided herein is a method for treating, preventing,or ameliorating one or more symptoms of diabetes or neurodegeneration ina subject, comprising administering to the subject a therapeuticallyeffective amount of a compound provided herein, e.g., a compound ofFormula I, or an enantiomer, a mixture of enantiomers, a mixture of twoor more diastereomers, or an isotopic variant thereof; or apharmaceutically acceptable salt, solvate, or hydrate thereof.

In yet another embodiment, provided herein is a method for treating,preventing, or ameliorating one or more symptoms of diabetes in asubject, comprising administering to the subject a therapeuticallyeffective amount of a compound provided herein, e.g., a compound ofFormula I, or an enantiomer, a mixture of enantiomers, a mixture of twoor more diastereomers, or an isotopic variant thereof; or apharmaceutically acceptable salt, solvate, or hydrate thereof. In oneembodiment, the diabetes is type 1. In one embodiment, the diabetes istype 2.

In yet another embodiment, provided herein is a method for increasingglucose tolerance in a subject, comprising administering to the subjecta therapeutically effective amount of a compound provided herein, e.g.,a compound of Formula I, or an enantiomer, a mixture of enantiomers, amixture of two or more diastereomers, or an isotopic variant thereof; ora pharmaceutically acceptable salt, solvate, or hydrate thereof.

In yet another embodiment, the disclosure provides methods for treatinga neurological or neurodegenerative disorder in a subject comprisingadministering to the subject an effective amount of one or morecompounds of formula I.

In yet another embodiment, the disclosure provides compositions fortreating a neurological or neurodegenerative disorder comprising one ormore compounds of formula I.

In some embodiments, the neurological or neurodegenerative disorder isAlzheimer's disease, Parkinson's disease, Huntington's disease,fronto-temporal dementia, Pick's disease, chronic traumaticencepholopathy, traumatic brain injury, stroke, cerebellar ataxia,multiple sclerosis, Down syndrome, or an aging-related CNS disorder. Insome embodiments, the neurological or neurodegenerative disorder isAlzheimer's disease.

In certain embodiments, the subject is a mammal. In certain embodiments,the subject is a human. In certain embodiments, the subject is a primateother than a human, a farm animal such as cattle, a sport animal, or apet such as a horse, dog, or cat.

In the treatment, prevention, or amelioration of one or more symptoms ofthe disorders, diseases, or conditions described herein, an appropriatedosage level generally is ranging from about 0.001 to 100 mg per kgsubject body weight per day (mg/kg per day), from about 0.01 to about 75mg/kg per day, from about 0.1 to about 50 mg/kg per day, from about 0.5to about 25 mg/kg per day, or from about 1 to about 20 mg/kg per day,which can be administered in single or multiple doses. Within thisrange, the dosage can be ranging from about 0.005 to about 0.05, fromabout 0.05 to about 0.5, from about 0.5 to about 5.0, from about 1 toabout 15, from about 1 to about 20, or from about 1 to about 50 mg/kgper day.

For oral administration, the pharmaceutical compositions provided hereincan be formulated in the form of tablets containing from about 1.0 toabout 1,000 mg of the active ingredient, in one embodiment, about 1,about 5, about 10, about 15, about 20, about 25, about 50, about 75,about 100, about 150, about 200, about 250, about 300, about 400, about500, about 600, about 750, about 800, about 900, and about 1,000 mg ofthe active ingredient for the symptomatic adjustment of the dosage tothe patient to be treated. The pharmaceutical compositions can beadministered on a regimen of 1 to 4 times per day, including once,twice, three times, and four times per day.

It will be understood, however, that the specific dose level andfrequency of dosage for any particular patient can be varied and willdepend upon a variety of factors including the activity of the specificcompound employed, the metabolic stability and length of action of thatcompound, the age, body weight, general health, sex, diet, mode and timeof administration, rate of excretion, drug combination, the severity ofthe particular condition, and the host undergoing therapy.

In one embodiment, provided herein is a method for reducing stress in anER, comprising contacting the ER with an effective amount of a compoundprovided herein, e.g., a compound of Formula I, or an enantiomer, amixture of enantiomers, a mixture of two or more diastereomers, or anisotopic variant thereof; or a pharmaceutically acceptable salt,solvate, or hydrate thereof. In one embodiment, the ER stress isresulted from the perturbation of ER Ca²⁺ homeostasis.

In yet another embodiment, provided herein is a method for restoring ormaintaining calcium ion homeostasis in an ER, comprising contacting theER with an effective amount of a compound provided herein, e.g., acompound of Formula I, or an enantiomer, a mixture of enantiomers, amixture of two or more diastereomers, or an isotopic variant thereof, ora pharmaceutically acceptable salt, solvate, or hydrate thereof.

In yet another embodiment, provided herein is a method for increasingthe Ca²⁺ concentration of an ER, comprising contacting the ER with aneffective amount of a compound provided herein, e.g., a compound ofFormula I, or an enantiomer, a mixture of enantiomers, a mixture of twoor more diastereomers, or an isotopic variant thereof; or apharmaceutically acceptable salt, solvate, or hydrate thereof.

In yet another embodiment, provided herein is a method for modulatingthe activity of a SERCA, comprising contacting the SERCA with aneffective amount of a compound provided herein, e.g., a compound ofFormula I, or an enantiomer, a mixture of enantiomers, a mixture of twoor more diastereomers, or an isotopic variant thereof; or apharmaceutically acceptable salt, solvate, or hydrate thereof.

In certain embodiments, the SERCA is SERCA1. In certain embodiments, theSERCA is SERCA2. In certain embodiments, the SERCA is SERCA3.

In certain embodiments, the SERCA is SERCA1a. In certain embodiments,the SERCA is SERCA1b. In certain embodiments, the SERCA is SERCA2a. Incertain embodiments, the SERCA is SERCA2b. In certain embodiments, theSERCA is SERCA3a. In certain embodiments, the SERCA is SERCA3b. Incertain embodiments, the SERCA is SERCA3c.

The compound provided herein, e.g., a compound of Formula I, or anenantiomer, a mixture of enantiomers, a mixture of two or morediastereomers, or an isotopic variant thereof; or a pharmaceuticallyacceptable salt, solvate, or hydrate thereof; can also be combined orused in combination with other agents or therapies useful in thetreatment, prevention, or amelioration of one or more symptoms of thedisorders, diseases, or conditions for which the compounds providedherein are useful.

Suitable other therapeutic agents can also include, but are not limitedto, (1) alpha-adrenergic agents; (2) antiarrhythmic agents; (3)anti-atherosclerotic agents, such as ACAT inhibitors; (4) antibiotics,such as anthracyclines, bleomycins, mitomycin, dactinomycin, andplicamycin; (5) anticancer agents and cytotoxic agents, e.g., alkylatingagents, such as nitrogen mustards, alkyl sulfonates, nitrosoureas,ethylenimines, and triazenes; (6) anticoagulants, such as acenocoumarol,argatroban, bivalirudin, lepirudin, fondaparinux, heparin, phenindione,warfarin, and ximelagatran; (7) anti-diabetic agents, such as biguanides(e.g., metformin), glucosidase inhibitors (e.g., acarbose), insulins,meglitinides (e.g., repaglinide), sulfonylureas (e.g., glimepiride,glyburide, and glipizide), thiozolidinediones (e.g., troglitazone,rosiglitazone, and pioglitazone), and PPAR-gamma agonists; (8)antifungal agents, such as amorolfine, amphotericin B, anidulafungin,bifonazole, butenafine, butoconazole, caspofungin, ciclopirox,clotrimazole, econazole, fenticonazole, filipin, fluconazole,isoconazole, itraconazole, ketoconazole, micafungin, miconazole,naftifine, natamycin, nystatin, oxyconazole, ravuconazole, posaconazole,rimocidin, sertaconazole, sulconazole, terbinafine, terconazole,tioconazole, and voriconazole; (9) antiinflammatories, e.g.,non-steroidal anti-inflammatory agents, such as aceclofenac, acemetacin,amoxiprin, aspirin, azapropazone, benorilate, bromfenac, carprofen,celecoxib, choline magnesium salicylate, diclofenac, diflunisal,etodolac, etoricoxib, faislamine, fenbufen, fenoprofen, flurbiprofen,ibuprofen, indometacin, ketoprofen, ketorolac, lornoxicam, loxoprofen,lumiracoxib, meclofenamic acid, mefenamic acid, meloxicam, metamizole,methyl salicylate, magnesium salicylate, nabumetone, naproxen,nimesulide, oxyphenbutazone, parecoxib, phenylbutazone, piroxicam,salicyl salicylate, sulindac, sulfinpyrazone, suprofen, tenoxicam,tiaprofenic acid, and tolmetin; (10) antimetabolites, such as folateantagonists, purine analogues, and pyrimidine analogues; (11)anti-platelet agents, such as GPIIb/IIIa blockers (e.g., abciximab,eptifibatide, and tirofiban), P2Y(AC) antagonists (e.g., clopidogrel,ticlopidine and CS-747), cilostazol, dipyridamole, and aspirin; (12)antiproliferatives, such as methotrexate, FK506 (tacrolimus), andmycophenolate mofetil; (13) anti-TNF antibodies or soluble TNF receptor,such as etanercept, rapamycin, and leflunimide; (14) aP2 inhibitors;(15) beta-adrenergic agents, such as carvedilol and metoprolol; (16)bile acid sequestrants, such as questran; (17) calcium channel blockers,such as amlodipine besylate; (18) chemotherapeutic agents; (19)cyclooxygenase-2 (COX-2) inhibitors, such as celecoxib and rofecoxib;(20) cyclosporins; (21) cytotoxic drugs, such as azathioprine andcyclophosphamide; (22) diuretics, such as chlorothiazide,hydrochlorothiazide, flumethiazide, hydroflumethiazide,bendroflumethiazide, methylchlorothiazide, trichloromethiazide,polythiazide, benzothiazide, ethacrynic acid, ticrynafen,chlorthalidone, furosenide, muzolimine, bumetanide, triamterene,amiloride, and spironolactone; (23) endothelin converting enzyme (ECE)inhibitors, such as phosphoramidon; (24) enzymes, such asL-asparaginase; (25) Factor VIIa Inhibitors and Factor Xa Inhibitors;(26) farnesyl-protein transferase inhibitors; (27) fibrates; (28) growthfactor inhibitors, such as modulators of PDGF activity; (29) growthhormone secretagogues; (30) HMG CoA reductase inhibitors, such aspravastatin, lovastatin, atorvastatin, simvastatin, NK-104 (a.k.a.itavastatin, nisvastatin, or nisbastatin), and ZD-4522 (also known asrosuvastatin, atavastatin, or visastatin); neutral endopeptidase (NEP)inhibitors; (31) hormonal agents, such as glucocorticoids (e.g.,cortisone), estrogens/antiestrogens, androgens/antiandrogens,progestins, and luteinizing hormone-releasing hormone antagonists, andoctreotide acetate; (32) immunosuppressants; (33) mineralocorticoidreceptor antagonists, such as spironolactone and eplerenone; (34)microtubule-disruptor agents, such as ecteinascidins; (35)microtubule-stabilizing agents, such as pacitaxel, docetaxel, andepothilones A-F; (36) MTP Inhibitors; (37) niacin; (38)phosphodiesterase inhibitors, such as PDE III inhibitors (e.g.,cilostazol) and PDE V inhibitors (e.g., sildenafil, tadalafil, andvardenafil); (39) plant-derived products, such as vinca alkaloids,epipodophyllotoxins, and taxanes; (40) platelet activating factor (PAF)antagonists; (41) platinum coordination complexes, such as cisplatin,satraplatin, and carboplatin; (42) potassium channel openers; (43)prenyl-protein transferase inhibitors; (44) protein tyrosine kinaseinhibitors; (45) renin inhibitors; (46) squalene synthetase inhibitors;(47) steroids, such as aldosterone, beclometasone, betamethasone,deoxycorticosterone acetate, fludrocortisone, hydrocortisone (cortisol),prednisolone, prednisone, methylprednisolone, dexamethasone, andtriamcinolone; (48) TNF-alpha inhibitors, such as tenidap; (49) thrombininhibitors, such as hirudin; (50) thrombolytic agents, such asanistreplase, reteplase, tenecteplase, tissue plasminogen activator(tPA), recombinant tPA, streptokinase, urokinase, prourokinase, andanisoylated plasminogen streptokinase activator complex (APSAC); (51)thromboxane receptor antagonists, such as ifetroban; (52) topoisomeraseinhibitors; (53) vasopeptidase inhibitors (dual NEP-ACE inhibitors),such as omapatrilat and gemopatrilat; and (54) other miscellaneousagents, such as, hydroxyurea, procarbazine, mitotane,hexamethylmelamine, and gold compounds.

In certain embodiments, the other therapies that may be used incombination with the compounds provided herein include, but are notlimited to, surgery, endocrine therapy, biologic response modifiers(e.g., interferons, interleukins, and tumor necrosis factor (TNF)),hyperthermia and cryotherapy, and agents to attenuate any adverseeffects (e.g., antiemetics).

Such other agents, or drugs, can be administered, by a route and in anamount commonly used therefor, simultaneously or sequentially with thecompound provided herein, e.g., a compound of Formula I, or anenantiomer, a mixture of enantiomers, a mixture of two or morediastereomers, or an isotopic variant thereof, or a pharmaceuticallyacceptable salt, solvate, hydrate, or prodrug thereof. When a compoundprovided herein is used contemporaneously with one or more other drugs,a pharmaceutical composition containing such other drugs in addition tothe compound provided herein can be utilized, but is not required.Accordingly, the pharmaceutical compositions provided herein includethose that also contain one or more other active ingredients ortherapeutic agents, in addition to a compound provided herein.

6. EXAMPLES

The disclosure will be further understood by the following non-limitingexamples.

As used herein, the symbols and conventions used in these processes,schemes and examples, regardless of whether a particular abbreviation isspecifically defined, are consistent with those used in the contemporaryscientific literature, for example, the Journal of the American ChemicalSociety or the Journal of Biological Chemistry. Specifically, butwithout limitation, the following abbreviations may be used in theexamples and throughout the specification: g (grams); mg (milligrams);mL (milliliters); μL (microliters); M (molar); mM (millimolar); μM(micromolar); mol (moles); mmol (millimoles); hr or hrs (hour or hours);and min (minutes).

For all of the following examples, standard procedures and methods knownto those skilled in the art can be utilized. Unless otherwise indicated,all temperatures are expressed in ° C. (degrees Centigrade). Allprocedures are conducted at room temperature unless otherwise noted.

6.1. Biological Assays

6.1.1. Cell Survival Assay

CSM14.1 neuronal cells were maintained in Complete Medium at 32° C.;wherein the Complete Medium contained Dulbecco's modified eagle medium(DMEM) with 10% fetal bovine serum (FSB), 1% L-glutamine, 100 IU/mLpenicillin, and 100 μg/mL streptomycin. The cells were recovered fromcultures by trypsinization and then seeded in 384 well plates (Greiner#781098) at a concentration of 1,000 cells/well in 20 μL of DMEM AssayMedium, wherein the DMEM Assay Medium contained 2% FBS, 100 IU/mLpenicillin, and 100 μg/mL streptomycin. Seeding was performed using aMultiDrop Combi reagent broadcaster. The plates were incubated overnightat 32° C. A test compound was prepared by 2-fold serial dilution in 100%DMSO using a BIOMEK® 2000 liquid handler (Beckman Coulter). Adose-response curve containing 10 concentrations of the test compoundwas obtained. Using a BIOMEK® FX liquid handler (Beckman Coulter), 2.5μL of the test compound was transferred from the 100% DMSO serialdilution plate to an intermediate plate containing 47.5 μL of DMEM AssayMedium containing 2% FBS, 100 IU/mL penicillin, and 100 μg/mLstreptomycin and mixed. To reduce or eliminate the interference from thecompound's precipitation, 6 μL of the diluted compound was immediatelytransferred to the assay plate to achieve a high compound concentrationof 100 μM in 99% DMEM Assay Medium and 1% DMSO. After the assay plateswere incubated for 2 hrs, 4 μL of 112.5 μM thapsigargin (TG) (DMSO stockdiluted into Assay TC Medium) was dispensed into each testing well witha MultiDrop Combi reagent broadcaster for a final concentration of about15 μM TG. The tissue culture media (4 μL) containing vehicle only wastransferred manually to each control cell using a 16 channel electronicpipette. After the plates were incubated overnight (about 16 to 24 hrs),CELLTITER-GLO® (Promega) (16 μL) was added to all wells and luminescencewas measured. High luminescence indicates cell survival. The biologicalresults are summarized in Table 1 where A represents a cell viabilityEC₅₀ less than 1 μM and B represents a value greater than 1 μM and lessthan 10 μM.

TABLE 1 Cmpd. Cell Survival EC₅₀ B1 B B2 B B3 B B4 A

6.1.2. Cell Rescue Assays

Protection against Thapsigargin (TG)-induced Cell-death. Human embryonickidney (HEK293) cells were grown in Dulbecco's Modified Eagle's Medium(DMEM) with 10% FBS and 1% antibiotic antimycotic solution (ABAM). Mouseneuroblastoma (N2a) cells were grown in 1:1 DMEM:OPTI-MEM® with 5% FBSand 1% ABAM. All cells were grown in 10% CO₂ in a humidified environmentof the incubator. Cells grown in 96-well plates were exposed to a testcompound (20 μM) for 2 hrs before addition of thapsigargin (15 μM forHEK293 cells and 1 μM for N2a cells) to induce ER stress. Afterincubation in cell-culture incubator for 24 hrs, ALAMARBLUE® regent (10%v/v) was added to the wells. Fluorescence reading was taken 2 hrs afterthe addition of the ALAMARBLUE® regent. Cell-viability was calculated asa percentage of relative fluorescence unit (RFU) compared to control.Vehicle-treated control cells showed similar viability as that ofuntreated cells. The biological results are summarized in Table 2, whereeach compound was tested at 20 μM and where A represents a viabilitygreater than 50%, B represents a viability greather than 10% and lessthan 50%, C represents a value below 10%.

TABLE 2 Cmpd. Cell Survival % B1 B B2 B B3 A B4 A

Protection against ß-amyloid-induced cell toxicity. Chonpathompikunlertet al. Eu. J. Pharmacol. 2011, 650, 544-549. Human SH-SY5Y cells aresuspended in media containing 2% FBS and 2× Gentamicin. Transfer 100ul/well culture to 96-well plate (20,000 cells/well; passage #5) andincubate overnight. Prepare compound samples at 30 uM. Add 50 ul of thesamples to the cells [concentration of compounds 10 uM] and incubate 3hours. Make stock of 40 uM AB1-42 peptide in assay media. Transfer 50ul/well to cells/compounds in assay plate. The final assay volume is 200ul/well, containing 2% FBS, 2× Gentamicin, 7.5 uM compound and AB1-42 at10 uM. Incubate for 4 days then add 20 ul of Promega Substrate CellTiter 96 Aqueous One Solution Reagent to each well. Incubate 37° C. andread OD at 490 nm. The results are summarized in Table 3, where Arepresents a value greater than 50% cell rescue, B represents a valuegreater than 10% and less than 50% cell rescue, and C represents a valuegreater than 1% and less than 10% cell rescue.

TABLE 3 Cmpd. Cell Survival % B1 B B2 A B3 C B4 A

6.1.3. Ca-ATPase Assay

A Ca-ATPase assay was performed using microsomal preparations from HEK293 cells at a series of calcium concentrations corresponding to thephysiological range, relative to controls. The ATP hydrolysis rate wasmeasured over a range of calcium concentrations in the presence of testcompounds using an NADH-linked, enzyme-coupled ATPase assay adapted for96-well microplates, with V_(max) determined by fitting the ATPasecalcium-dependence to the Hill function. Each well contained 2 μg or 7μg of SR vesicles (optimized for skeletal or cardiac SR, respectively),50 mM MOPS (pH 7.0), 100 mM KCl, 5 mM MgCl₂, 1 mM EGTA, 0.2 mM NADH, 1mM phosphoenol pyruvate, 5 IU pyruvate kinase, 5 IU lactatedehydrogenase, and 3.5 μg/mL A23187 (a calcium ionophore). CaCl₂ wasadded to set free [Ca²⁺] to the specific values. The assay was startedupon addition of ATP at a final concentration of 5 mM, and read in aSpectraMax Plus microplate spectrophotometer. The results are summarizedin Table 4, where A represents an increase in SERCA Vmax greater than50% and B represents a value greater than 10% and less than 50%.

TABLE 4 SERCA Vmax Cmpd. Increase B1 A B2 B B3 C B4 A

6.1.4. Determination of the Effect of a SERCA Agonist on the Level ofBlood Glucose

Ob/ob mice (10 weeks old, n=7) were intraperitoneally (i.p.) injectedwith 100 μL of a solution containing 0 (vehicle) or 50 mg/kg of a testcompound once a day for a total of 5 days. Fasting glucose was measuredat baseline and 4 days after the administration of the test compound.Glucose levels were measured in blood samples drawn from the tail veinusing a handheld glucose sensor. Compound effects on blood glucose areshown in Table 5 where A represents a 10-20% reduction in blood glucose,B represents a 21-30% reduction in blood glucose, and C represents agreater than 30% reduction in blood glucose levels.

TABLE 5 Glucose level Cmpd. reduction B1 B B2 B B3 A B4 C

6.2. Compound Synthesis 6.2.1.3-(4-Bromothienyl-2-yl)-5-(2-methylquinolin-8-yl)-1,2,4-oxadiazole B3

3-(4-Bromothienyl-2-yl)-5-(2-methylquinolin-8-yl)-1,2,4-oxadiazole B3was synthesized as shown in Scheme 2.

A mixture of 4-bromothiophene-2-carbaldehyde, NH₂OH.HCl, and K₂CO₃ inEtOH was refluxed overnight to form an oxime. The oxime was thendissolved in SOCl₂ and the reaction mixture was stirred at roomtemperature for 2 hrs to yield crude 4-bromothiophene-2-carbonitrile.The crude 4-bromothiophene-2-carbonitrile was treated with NH₂OH.HCl inthe presence of Et₃N in EtOH at reflux overnight to form4-bromo-N-hydroxythiophene-2-carboximidamide. The reaction was completeas followed by LC/MS. The 4-bromo-N-hydroxythiophene-2-carboximidamide(1 eq.) was coupled subsequently with 2-methylquinoline-8-carboxylicacid (1 eq.) in the presence of CDI (1.2 eq.) and Et₃N (1 eq.) in DMF.The coupling was monitored by LC/MS. After the reaction was complete,acetic acid was charged and the mixture was fluxed overnight. After thereaction was complete, the desired compound,3-(4-bromothienyl-2-yl)-5-(2-methylquinolin-8-yl)-1,2,4-oxadiazole B3(100 mg), was purified via TLC and obtained in 30% yield.

6.2.2. 3-(4-Isopropoxyphenyl)-5-(2-methylquinolin-8-yl)-1,2,4-oxadiazoleB1

3-(4-Isopropoxyphenyl)-5-(2-methylquinolin-8-yl)-1,2,4-oxadiazole B1 wassynthesized according to the procedure as described for compound B3.4-Isopropoxy-benzaldehyde (1.0 mmol), hydroxylamine hydrochloride (1.5mmol), and potassium carbonate (2.0 mmol) were dissolved in anhydrousethanol and heated to reflux overnight. After cooling, the mixture wasdiluted with water and extracted 3 times with ethyl acetate. The oximewas isolated via evaporation of organic solvents. The residue wasdissolved in thionyl chloride (5 mL) and stirred at room temperature for2 hrs. After evaporation of excess thionyl chloride, the crude nitrilewas obtained. This product was dissolved in ethanol and treated with 3equivalents each of hydroxylamine hydrochloride and triethylamine, andheated to reflux overnight. This resulting oxime was isolated byextraction into ethyl acetate, and subsequently evaporated. The oximewas dissolved in dimethylformamide and treated with 1.5 equivalents ofcarbonyl diimidazole, 1.5 equivalents of triethylamine, and 1.2equivalents of 2-methylquinoline-8-carboxylic acid. After 5 hrs, aceticacid (2 mL) was added and the mixture was heated at reflux overnight.Evaporation of solvents afforded a residue that was purified bypreparative reverse-phase HPLC using a water-acetonitrile gradient toafford the desired3-(4-isopropoxyphenyl)-5-(2-methylquinolin-8-yl)-1,2,4-oxadiazole B1.

6.2.3.3-(5-Bromothiophen-2-yl)-5-(2-methylquinolin-8-yl)-1,2,4-oxadiazole B2

3-(5-Bromothiophen-2-yl)-5-(2-methylquinolin-8-yl)-1,2,4-oxadiazole B2was synthesized according to the procedure as described for compound B3.5-Bromo-2-formylthiophene (1.0 mmol), hydroxylamine hydrochloride (1.5mmol), and potassium carbonate (2.0 mmol) were dissolved in anhydrousethanol and heated to reflux overnight. After cooling, the mixture wasdiluted with water and extracted 3 times with ethyl acetate. The oximewas isolated via evaporation of organic solvents, and the residue wasdissolved in thionyl chloride (5 mL), and stirred at room temperaturefor 2 hrs. After evaporation of excess thionyl chloride, the crudenitrile was obtained. This product was dissolved in ethanol and treatedwith 3 equivalents each of hydroxylamine hydrochloride and triethylamineand heated to reflux overnight. This resulting oxime was isolated byextraction into ethyl acetate, and subsequently evaporated. The oximewas dissolved in dimethylformamide and treated with 1.5 equivalents ofcarbonyl diimidazole, 1.5 equivalents of triethylamine, and 1.2equivalents of 2-methylquinoline-8-carboxylic acid. After 5 hrs, aceticacid (2 mL) was added and the mixture was heated at reflux overnight.Evaporation of solvents afforded a residue that was purified bypreparative reverse-phase HPLC using a water-acetonitrile gradient toafford the desired3-(5-bromothiophen-2-yl)-5-(2-methylquinolin-8-yl)-1,2,4-oxadiazole B2.

6.2.4. 3-(4-Chlorophenyl)-5-(2-methylquinolin-8-yl)-1,2,4-oxadiazole B4

3-(4-Chlorophenyl)-5-(2-methylquinolin-8-yl)-1,2,4-oxadiazole B4 wassynthesized according to the procedure as described for compound B3.4-Chlorobenzaldehyde (1.0 mmol), hydroxylamine hydrochloride (1.5 mmol),and potassium carbonate (2.0 mmol) were dissolved in anhydrous ethanoland heated to reflux overnight. After cooling, the mixture was dilutedwith water and extracted 3 times with ethyl acetate. The oxime wasisolated via evaporation of organic solvents. The residue was dissolvedin thionyl chloride (5 mL) and stirred at room temperature for 2 hrs.After evaporation of excess thionyl chloride, the crude nitrile wasobtained. This product was dissolved in ethanol and treated with 3equivalents each of hydroxylamine hydrochloride and triethylamine, andheated to reflux overnight. This resulting oxime was isolated byextraction into ethyl acetate, and subsequently evaporated. The oximewas dissolved in dimethylformamide and treated with 1.5 equivalents ofcarbonyl diimidazole, 1.5 equivalents of triethylamine, and 1.2equivalents of 2-methylquinoline-8-carboxylic acid. After 5 hrs, aceticacid (2 mL) was added and the mixture was heated at reflux overnight.Evaporation of solvents afforded a residue that was purified bypreparative reverse-phase HPLC using a water-acetonitrile gradient toafford the desired3-(4-chlorophenyl)-5-(2-methylquinolin-8-yl)-1,2,4-oxadiazole B4.

The examples set forth above are provided to give those of ordinaryskill in the art with a complete disclosure and description of how tomake and use the claimed embodiments, and are not intended to limit thescope of what is disclosed herein. Modifications that are obvious topersons of skill in the art are intended to be within the scope of thefollowing claims. All publications, patents, and patent applicationscited in this specification are incorporated herein by reference as ifeach such publication, patent or patent application were specificallyand individually indicated to be incorporated herein by reference.

What is claimed:
 1. A compound of Formula I:

or an enantiomer, a mixture of enantiomers, a mixture of two or morediastereomers, or an isotopic variant thereof; or a pharmaceuticallyacceptable salt, solvate, or hydrate thereof; wherein: R² is methyl; R³,R⁴, R⁵, R⁶, and R⁷ are each independently (a) hydrogen, cyano, or haloor (b) C₁₋₆ alkyl with the proviso that R³ and R⁷ are not concurrentlyCl; and R¹ is optionally substituted phenyl, thiophenyl, or furanyl withthe proviso that R₁ is not 2-methoxy-phenyl or 2-butoxy-phenyl.
 2. Thecompound according to claim 1, or a pharmaceutically acceptable saltthereof, wherein R³, R⁴, R⁵, R⁶, and R⁷ are hydrogen.
 3. The compoundaccording to claim 1, or a pharmaceutically acceptable salt thereof,wherein R¹ is mono or disubstituted phenyl or 2-thienyl.
 4. The compoundaccording to claim 1 selected from the group consisting of: